★产品分类导航
产品英文名:Fluoro-Gold
产品中文名: 荧光金
产品编号:471905      产品品牌:fluorochrome

价格:内详

  1. Fluoro-Gold
  2. Use Guide and Protocol
  3. US Patent No. 4716905
  4. UK Patent No. 2181545

Fluoro-Gold

规格

价格

20 mg 3310元
50 mg 7910元
100 mg 14700元
150 mg 19600元
200 mg 23100元


美国fluorochrome471905,荧光金,Fluoro-Gold原装现货
Amygdala cells (40X) after Fluoro-Gold
injection in the PVN. Antibody is at 1/50''000.

Main Protocol

1. Background
The use of Fluoro-Gold is essentially the same as other fluorescent tracers. The main difference is that Fluoro-Gold is more flexible in terms of post-injection survival times'' concentration range'' tissue treatment and compatibility with other histochemical techniques.

2. Storage and Shelf Life
Dry Fluoro-Gold should be kept in a light tight closed container at 4 degrees Celsius. Stored properly'' Fluorogold should have a shelf life exceeding one year. The dye in solution should also be kept in a light tight closed container at 4 degrees Celsius and should remain stable for at least six months.

3. Vehicle
Fluoro-Gold can be dissolved in distilled water or 0.9% saline'' or utilized as a suspension
in 0.2M neutral phosphate buffer.

4. Dye Concentration
Fluoro-Gold has been successfully used at concentrations ranging from 1-10%. Initially'' a 4% concentration is advised. If undesirable necrosis occurs at the injection site'' or labeling is too intense'' reduce the concentration to a 2% solution. If you need to use more precise measurements'' the molecular weight of Fluoro-Gold is 532.6 daltons.

5. Dye Administration

A. Pressure Injection - This is probably the most frequently used mode of application. Volumes injected range from .05-1 μl'' typically .1-.2 μl.

B. Iontophoresis - Discrete'' small injection sites result from 4-10 second pulsed iontophoretic (+5 to +10ua/10min) application.

C. Crystal - A crystal of the tracer can be administered from the tip of a micro-pipette.

6. Post-0perative Survival Period
Good retrograde labeling has been observed with periods ranging from two days to two months. Survival periods of three to five days are typical. Long survival periods enhance filling of distal processes without diffusion of the dye from the cell.

7. Fixation
Almost any fixative'' or no fixative'' can be used'' Phosphate neutral buffered saline containing 4% formaldehyde is frequently employed. Fixatives containing high concentrations of heavy metals (e.g. osmium'' mercury) will quench the fluorescence'' while high concentrations (over 1%) of glutaraldehyde may increase background fluorescence

8. Histochemical Processing
Tissue containing Fluoro-Gold may be processed according to virtually any common histological technique. This includes cryostat sections of unfixed tissue (10 μm)'' frozen sections of fixed tissue (20 μm)'' and thin sections cut from tissue imbedded in either plastic (.2-4 μm) or paraffin (3-10 μm). Frozen sections of fixed tissue are most frequently used.

9. Combined Methods
At this point of processing'' sections may be further processed for a second marker such as autoradiography'' HRP histochemistry'' immunocytochemistry'' a second fluorescent tracer'' fluorescent counterstain'' etc.

10. Mounting'' Clearing and Coverslipping
Sections are typically mounted on gelatin-coated slides'' air-dried'' immersed in xylene'' and coverslipped with nonfluorescent DPX plastic mounting media. Sections may be dehydrated with graded alcohols'' unless this is not compatible with a second tracer. If Fluoro-Gold is to be combined with fluorescence immunocytochemistry'' then sections are air-dried and directly coverslipped with neutral buffered glycerine (1:2).

11. Examination and Photography
Fluoro-Gold can be visualized with a fluorescence microscope using a wide band ultraviolet excitation filter. A gold color is emitted when tissue has been processed with neutral pH buffer'' whereas a blue color is emitted when tissue is processed with acidic (e.g. pH 3.3) pH buffer. It can be photographed digitally or with film (use Ektachrome 200-400 ASA film for color prints and comparable speed film for black and white prints'' for example Tri-X). Most exposure times range from 10-60 second exposures'' depending on the objective magnification and the intensity of the label. Thirty (30) second exposures are about average. Multiple exposures may be exploited to simultaneously visualize Fluoro-Gold and another tracer. Thus'' UV would be combined with bright field illumination to simultaneously locate Fluoro-Gold with HRP or silver grains in autoradiography. Similarly'' blue light excitation can be combined to also visualize the green emission color of FITC'' while green excitation light may be used to simultaneously observe the red emission color of propidium iodide'' or ethidium bromide (a fluorescent counterstain).

Additional Information Concerning the Use of Fluoro-Gold

美国fluorochrome471905,荧光金,Fluoro-Gold原装现货Vehicle
For pressure injections through a microsyringe or micropipette'' Fluoro-Gold should be dissolved in distilled water or .9% saline. Fluoro-Gold may also be utilized as a suspension in .2M neutral phosphate buffer'' however'' the suspended particles may clog a fine micropipette tip so distilled water or .9% saline is the preferred vehicle. For iontophoresis'' a 1% Fluoro-Gold solution is made up in .1M acetate buffer (pH=3.3). Well-cleaned (95% ETOH'' water) glass micropipettes should have tips of 10-20 μm. Optimal iontophoresis parameters are +1 to +5u amps delivered with pulsed current (4-10 seconds on'' 4-10 seconds off) over a 10-20 minute period.

Injection Sites
Virtually any central or peripheral nervous system structure can be injected with Fluoro-Gold for analysis of retrograde transport. In the peripheral nervous system'' ganglia and peripheral targets can be studied. For studies of peripheral nerve'' the nerve should be cut or damaged and either dipped in'' or injected with'' aq 5% solution of Fluoro-Gold. Since Fluoro-Gold is not significantly taken up by intact fibers of passage'' the fibers must be cut or severely damaged for uptake of the dye to occur.

Transport and Survival Time
Fluoro-Gold is used as a retrograde axonal tracer'' although orthograde axonal transport does occur. The survival time should be varied (especially to very short survival times of 12 hours - 2 days) to maximize orthograde transport in the specific neuronal system under study. For retrograde transport'' the survival times should be varied from 4 days to 14 days. Seven to 10 days works for most systems'' although long pathways (e.g.'' spinal cord to brainstem) and pathways in large mammals (e.g.'' cats'' monkeys) may require longer survival times (e.g.'' 14 days). In addition'' since Fluoro-Gold remains fast within retrogradely labeled neurons'' survival times of several months will also produce excellent results. For iontophoresis'' a 2-5 day survival time is recommended. It is estimated that transport occurs at about 2 cm per day for mammals; it is slower for cold-blooded animals.

Tissue Processing
Tissue processing is covered in detail in the use guide and in the original publication (Schmued and Fallon'' 1986'' Brain Research 377:147-154). Since Fluoro-Gold is stable in many solvents and remains fast within retrogradely labeled neurons'' it’s use is compatible with many histochemical techniques. It can be used with other retrograde tracers'' immunofluorescence'' PAP and ABC immunocytochemistry'' HRP histochemistry'' autoradiography'' counterstains (ethidium bromide is the preferred fluorescent counterstain)'' paraffin embedding and plastic embedding. However'' if tissue is unfixed'' additional processing of tissue in aqueous solutions for over an hour or two will result in loss of Fluoro-Gold fluorescence from labeled neurons. Fluoro-Gold may be useful in electron microscopy. Fluoro-Gold can be used in a brain which has been sectioned and transferred to phosphate buffer. Sections are typically mounted on gelatin-coated slides'' air dried'' immersed in xylene and coverslipped with DPX plastic mounting media (FLUKA Chemical Corp.'' 255 Oser Avenue'' Hauppauge'' New York'' 11788'' Catalog #44581). Tissue may also be viewed on slides without further processing'' can be run through graded alcohols for dehydration'' or'' for immunocytochemistry'' the sections can be air dried and directly coverslipped with neutral buffered glycerine (1:2).

Examination and Photography
Fluoro-Gold is visualized with a fluorescence microscope using a wide band ultraviolet (UV) excitation filter. Use the same filter pack you would for other fluorescent retrograde tracers excited under wide band UV (e.g.'' True Blue'' Fast Blue'' Nuclear Yellow)'' such as the Leitz Ploem filter system A (Wide Band UV'' Excitation filter BP 340-380)'' Mirror RKP 400'' Barrier Filter LP 430). Objectives should be made especially for fluorescence microscopy (such as that made by Zeiss) glycerine'' or water. Since plastic does absorb UV light'' it is not advised to view through plastic petri dishes'' etc. Recommended films are T-Max (Kodak'' black & white) and Ektachrome 200 (Kodak'' color slides). Exposure times usually vary from 20 seconds to 1.5 minutes.

Chemical Analysis

Quality Expected Result Actual Result
Appearance A golden-yellow'' hygroscopic'' crystalline powder A bright-yellow powder
Odor None None
Solution 20 ml of a 5% w/v aqueous solution should be clean'' clear and almost free from suspended matter'' and should have not more than a very slight odor Passes Test
pH of a 1% Solution Between 4.0 and 5.5 at 25 degrees Celsius 4.6
Spectral characteristics The spectral characteristics of Fluoro-Gold vary with pH A 0.1% solution in distilled water has a pH of 4.5 and excitation peak of 414 nm and emission peak of 541 nm

Fluoro-Gold bound to membranes at a physiological pH of 7.4 has an excitation band of 350 to 395 nm and an emission band of 530 to 600 nm

Chloride Not more than 0.035% 0.017%
Sulfate Not more than 0.1% Less than 0.05%
Sulfated ash Not more than 0.1% Negligible
Heavy metals Not more than 10 p.p.m Less than 10 p.p.m
Selenium Not more than 30 p.p.m Less than 10 p.p.m.
Loss on drying Not more than 1.0% after 3 hours in vacuo at 60 degrees Celsius 0.1%
Assay Between 95.0 and 105.0% calculated with reference to the dried material 99.2%

Notice: The original and only true Fluoro-Gold (Fluorogold) is produced by Fluorochrome'' LLC and marketed by Fluorochrome'' LLC and Histo-Chem Inc.

Fluoro-Gold (Fluorogold) is an exclusive product of Fluorochrome'' LLC. It has been sold by Fluorochrome and widely used since 1985. Other companies are marketing a product they claim is the same as or equivalent to Fluoro-Gold. In fact'' the chemical structures of these compounds seem to be different from Fluoro-Gold. Certain physical properties of the compounds may be very different.

*CAUTION: Fluoro-Gold'' Antibody to Fluoro-Gold and Fluoro-Ruby are for investigational use only in laboratory research animals or for tests in vitro. NOT FOR USE IN HUMANS. These drugs should be used only by persons regularly engaged in conducting neuroanatomical studies and tests in vitro or in animals used only for laboratory research.

fluorochrome471905,2181545,荧光金抗体产品

Fluorochrome公司是专业生产荧光金及荧光金抗体的专业高科技公司。公司拳头产品荧光金于1985年推向市场,广泛用于神经生物学研究。荧光金在紫外线激发下发金黄色光,特点是非常灵敏,不仅能标记胞浆,而且能很好地显示树突分枝,但核和核仁不染色;在胞体内分解慢,甚至在注射后存活2月标记强度仍无明显变化;比较耐紫外线的照射,褪色比较慢;可以经受许多组织学染色处理,因而可以和HRP、免疫组织化学等方法结合。Fluorochrome公司还提供荧光金抗体及红色荧光金,扩大了其应用范围

Major Publications

Balercia'' G.'' Chen'' S.'' and Bentivoglio'' M.'' "Electron microscopic analysis of fluorescent neuronal labeling after photoconversion''" Journal of Neuroscience Methods 45 (1992) 87-98.

*Pieribone'' V.A.'' Aston-Jones'' G.'' "The iontophoretic application of Flouro-Gold for the study of the afferents to deep brain nuclei''" Brain Research 475 (1988) 259-271.

Reep'' R.L.'' Baccala'' M.J.'' Booth'' M.P.'' and Goodwin'' G.S.'' "Combined retrograde and anterograde tracing of neuronal connections: Fluoro-Gold and autoradiography''" Journal of Neuroscience Methods'' 23 (1988) 1-5.

Schmued'' L.'' "Fluoro-Gold and 4-Acetamido-4’-isothiocyanostilbene-2''2’disulfonic Acid: Use of Substituted Stilbenes in Neuroanatomical Studies''" Methods in Neurosciences'' 3 (1990) 317-330.

*Schmued'' L.C. and Fallon'' J.H.'' "Fluoro-Gold: a fluorescent retrograde axonal tracer with numerous unique properties''" Brain Research'' 377 (1986) 147-154.

*Schmued'' L.C.'' "Anti-retrograde and retrograde neuroanatomical tract tracing with fluorescent compounds''" Neuroscience Protocols'' 94-050-02 (1994) 1-15.

Van Bockstaele'' E.J.'' Wright'' A.M.'' Cestari'' D.M.'' and Pickel'' V.M.'' "Immunolabeling of retrogradely transported Fluoro-Gold: sensitivity and application to ultrastructural analysis of transmitter-specific mesolimbic circuitry''" Journal of Neuroscience Methods'' 55 (1994) 65-78.

*Bowyer'' J.F.'' Gough'' B.'' Broening'' H.W.'' Newport'' G.D.'' and Schmued'' L.'' "Fluoro-Gold and Pentamidine Inhibit the In Vitro and In Vivo Release of Dopamine in the Striatum of Rat''" The Journal of Pharmacology and Experimental Therapeutics'' 266 (1993) 1066-1074.

*Chang'' H.T.'' Kuo'' H.'' Whittaker'' J.A.'' and Cooper'' N.G.F.'' "Light and electron microscopic analysis of projection neurons retrogradely labeled with Fluoro-Gold notes on the application of antibodies of Fluoro-Gold''" Journal of Neuroscience Methods'' 35 (1990) 31-37.

Schmued'' L.C.'' Kyriakidis'' K.'' Fallon'' J.H.'' and Ribak'' C.E.'' "Neurons containing retrogradely transported Fluoro-Gold exhibit a variety of iysosomal profiles: a combined brightfield'' fluorescence and electron microscopic study''" Journal of Neurocytology'' 18 (1989) 333-343.

Schmued'' L.'' Kyriakidis'' K.'' & Heimer'' L.'' In vivo anterograde and retrograde axonal transport of the fluorescent rhodamine-dextran-amine'' Fluoro-Ruby'' within the CNS. Brain Res'' 526 (1990) 127-134.

Bowyer'' J.F. and Schmued'' L.'' Fluoro-Ruby labeling prior to an amphetamine neurotoxic insult shows a definitive massive loss of dopaminergic terminals and axons in caudateputamen. Brain Res.'' 1075'' (2006) 236-239.

*KEY REFERENCES (See Medline database "Fluoro-Gold" for 1505 citations from July 1986 to November 2009'' plus an additional 850 citations for "fluorogold" as of november 2009)

Notice: The original and only true Fluoro-Gold (Fluorogold) is produced by Fluorochrome'' LLC and marketed by Fluorochrome'' LLC and Histo-Chem Inc.

Fluoro-Gold (Fluorogold) is an exclusive product of Fluorochrome'' LLC. It has been sold by Fluorochrome and widely used since 1985. Other companies are marketing a product they claim is the same as or equivalent to Fluoro-Gold. In fact'' the chemical structures of these compounds seem to be different from Fluoro-Gold. Certain physical properties of the compounds may be very different.

*CAUTION: Fluoro-Gold'' Antibody to Fluoro-Gold and Fluoro-Ruby are for investigational use only in laboratory research animals or for tests in vitro. NOT FOR USE IN HUMANS. These drugs should be used only by persons regularly engaged in conducting neuroanatomical studies and tests in vitro or in animals used only for laboratory research.

应用文献:

Antibodies

Fluorochrome sells the following high quality antibodies:

  • Antibody to Fluoro-Gold
  • Ki-67 Antibody-W
  • GFAP Antibody-W
  • BrdU Antibody-W

Our Antibody to Fluoro-Gold is by far the most effective antibody to

Fluoro-Gold available. Since Fluorochrome first introduced it in 1998, it has a achieved a proven record of accomplishment and has become the standard in the industry.

Ki-67 Antibody-W
*Includes blocking peptide.

3940元
Introductory Offer

GFAP Antibody-W
*Includes blocking peptide.

3940元
Introductory Offer

BrdU Antibody-W
*Includes blocking agent.

3940元



The Ki-67 Antibody-W, GFAP Antibody-W, and BrdU Antibody-W are excellent antibodies that are recent introductions. Included with these antibodies is filter paper absorbed with a specific blocking peptide. We feel that a blocking agent should be utilized in all investigations using these types of antibodies. Only with a blocking agent can the researcher verify and have confidence that the observed signal is the intended signal. At no additional costs, we have included the blocking agent, which makes this verification easy and accurate for the researcher to implement.

As an introductory offer, we are discounting the purchase price for the Ki-67 Antibody-W, GFAP Antibody-W, and BrdU Antibody-W by 20%.

Fluoro-Gold Antibody Use Guide and Protocol

美国fluorochrome471905,荧光金,Fluoro-Gold原装现货
Hypothalamic cells along the 3rd left
ventricle (40X) after Flouro-Gold injection
in the PVN. Antibody is at 1/50,000.

Storage and Preparation of the Antibody

You will receive the Antibody to Fluoro-Gold in a small vial. It is a polyclonal antibody raised in rabbit. The vial will contain approximately 100μl of the antibody solution. It should be immediately frozen and stored in a cold (-20 degrees C), dark, dry environment. The freezer section, without the frost free feature, of a good commercial refrigerator should suffice. If properly and continuously frozen, the antibody solution can be stored up to one year.

The antibody solution should remain frozen until ready for use. The 100μl aliquot of the Antibody is at a dilution of 1/100. The Antibody can be further diluted 1/50,000 to 1/100,000 times in BSA diluent (50 mM KPBS, 0.4% Triton, 1% BSA, 1% NGS) to produce 50ml to 100ml of working titer. This means you can dilute the solution you receive by 500 to 1,000 times. This should treat 300 to 1000 sections if you use the Vector elite kit. After preparation, the antibody solution can be stored for up to seven days in a cool (4 degrees C), dark and dry environment. The refrigerator section of a good commercial refrigerator should suffice. We do not recommend that the solution be used beyond seven days after preparation. Do not refreeze the antibody solution.

Use of the Antibody

Fluoro-Gold can be injected using several different methods, including pressure, iontophoretic and other applications developed by a variety of researchers. See Schmued and Fallon, Fluoro-Gold: "A fluorescent retrograde axonal tracer with numerous unique properties," Brain Research, 377 (1986) 147-154 as well as Pieribone and Aston-Jones, "The Iontophoretic Application of Fluoro-Gold for the study of afferents to deep brain nuclei," Brain Research, 475 (1988) 259-271. Many researchers have developed their own modified procedures. Use of the antibody should not be dependent upon the methodology used to employ Fluoro-Gold.

After the Fluoro-Gold has been injected, floating sections (we used thirty um sections from a rat perfused with 4% formaldehyde) are incubated with the Fluoro-Gold Antibody solution overnight at 4 degrees C. Sections are washed, then incubated in Biotinylated GAR (Vector Labs) at 1/1000 for 1 hour at room temperature and washed again. Sections are then incubated in Avidin/Biotin (Vector Labs) at 1/1000 for 1 hour, washed and transferred to Diaminobenzidine (.04%) and Nickel Chloride (2.5%) in 0.1 M NaAcetate with 0.06% H2O2 for six minutes. Sections are then washed, mounted, dried, dehydrated and cover slipped.

It has been our experience that if stored and prepared in the manner set out above, each vial of the antibody should treat 300 to 1000 thirty um sections of the albino rat brain (or similar sized animals) if you use the Vector elite kit. However, you should experiment with the concentration and procedures to determine which best fits your circumstance.


Ki-67 Antibody-W Use Guide and Protocol

General:
Ki-67 Antibody-W reacts against the Ki-67 protein. The Ki-67 protein is an intrinsic marker of ongoing cell proliferation and is present during all active phases of the cell cycle (G1, S, G2 and mitosis) and is absent from resting cells (G0). This means it is an excellent marker for determining the growth fraction of a given cell population. It is used extensively in diagnostic, research and drug discovery applications. Ki-67 Antibody-W is polyclonal and was raised in rabbit to the human peptide sequence –TPKEKAQALEDLAGFKELFQT – that was coupled to thyroglobulin by glutaraldehyde before injection into the rabbit. Our antibody has been used for immunocytochemistry in rat and human brain tissue. It will be sold with filter paper absorbed with the specific peptide for blocking. This allows the researcher to confirm the specific signal.

Characterization of the Ki-67 Antibody-W

美国fluorochrome471905,荧光金,Fluoro-Gold原装现货
A. Ki-67 positive cells in the hippocampus (10X) of a 7 day old rat
B. Ki-67 cells in hippocampus (60X) taken on a confocal microscope
C. Ki-67 block in hippocampus (10X)

Specificity was ascertained by performing a blocking study. Adjacent sections from the hippocampus of a 7 day old rat were incubated with the Ki-67 Antibody-W alone (A-10X) and (B-60X) or with the Ki-67 Antibody-W preincubated with Ki-67 peptide (C-10X). Note labeling in A and B and the absence of labeling in C, indicating specificity of the antibody binding.

Storage and Preparation:
The vial you will receive contains approximately 200μl of solution at a dilution of 1/100. The Ki-67Antibody-W can be further diluted to at least 1/20,000 in BSA diluent (50mM KPBS, 0.4% Triton, 1% BSA, 1% NGS).

The antibody should be stored at -20C. Also included is filter paper absorbed with the specific peptide for blocking. 500μl of the diluted antibody can be added to the tube to achieve a 10μM block. Incubate for at least 1 hour at room temperature (RT) before applying to tissue. The antibody blocks specifically with this peptide and does not block with other peptides absorbed on filter paper.

Protocol:
Fresh frozen 10-30 micron sections are fixed in 4% paraformaldehyde (FA) for 1 hour at RT, washed in phosphate buffered saline (PBS), and then treated in 10% Sodium Citrate for 40 minutes at 90C. The sections are allowed to cool for 20 minutes in the sodium citrate solution, washed in PBS and incubated with 0.1 to 0.3% H2O2 in PBS for 10 to 30 minutes. The tissue is again washed in PBS, then treated with an Avidin/Biotin blocking kit (Vector-cat #SP-2001), washed and incubated with BSA diluent for 1 hour at RT. The Ki67 antibody (1/20,000) is applied to the tissue overnight at RT. After washing in PBS the sections are incubated in Biotinylated GAR at 1/1000 (Vector labs-cat#BA1000) for 1hr at RT, washed and then incubated in Avidin/Biotin at 1/1000 (Vector labs-cat#PK6100) for 1hr at RT. After washing the signal is visualized with Diaminobenzidine (0.04%) in 0.1M Sodium Acetate with 0.06% H2O2 for six minutes (Nickel Chloride at 2.5% may also be added). Sections are then water washed (counterstained if desired), dehydrated and cover slipped.

Ki-67 Antibody-W also works with the protocol used to visualize the BRDU Antibody-W sold by Fluorochrome, LLC. However, 4% FA perfused tissue may be used instead of fresh frozen sections.

Publications:
Perez et al, The Journal of Neuroscience, May 13, 2009:29(19):6379-6387 Garcia-Fuster et al, in preparation: Individual differences in adult hippocampal neurogensis following experimenter administration of cocaine.



GFAP Antibody-W Use Guide and Protocol


General:
GFAP Antibody-W reacts against the Glial Fibrillary Acidic Protein (GFAP), which is a class-III intermediate filament and the main constitute of intermediate filaments in astrocytes and serves as a cell specific marker for mature glial cells. It is useful as a marker of neural stem cells and astrocytic cells, including many types of brain tumor derived from astrocytic cells, which express GFAP. GFAP Antibody-W is polyclonal and was raised in rabbit to the peptide sequence NAGFKETRASERAE (human, rat and mouse) coupled to thyroglobulin by glutaraldehyde before injection into the rabbit. Our antibody has been used for immunocytochemistry in rat and human brain tissue. It will also be sold with filter paper absorbed with the specific peptide for blocking. This allows the researcher to confirm the specific signal.

Characterization of the GFAP Antibody-W

美国fluorochrome471905,荧光金,Fluoro-Gold原装现货
A. GFAP glail cells in hippocampus (10X) of an adult rat
B. GFAP glail cells in hippocampus (60X) taken on a confocal microscope
C. GFAP block in hippocampus (10)

Specificity was ascertained by performing a blocking study. Adjacent sections from the hippocampus of an adult rat were incubated with the GFAP Antibody-W alone (A-10X) and (B-60X) or with GFAP Antibody-W preincubated with GFAP peptide (C-10X). Note labeling in A and B in glial cells and the absence of labeling in C, indicating specificity of the antibody binding.

Storage and Preparation:
The vial you will receive contains approximately200μl at a dilution of 1/100. The antibody can be further diluted to at least 1/30,000 in BSA diluent (50mM KPBS, 0.4% Triton, 1% BSA, 1% NGS).

The antibody should be stored at -20C. Also included is filter paper absorbed with the specific peptide for blocking. 500μl of the diluted antibody can be added to the tube to achieve a 10 μM block. Incubate for at least 1 hour at room temperature (RT) before applying to tissue. The antibody blocks specifically with this peptide and does not block with other peptides absorbed on filter paper.

Protocol:
Fresh frozen 10-30 micron sections are fixed in 4% paraformaldehyde (FA) for 1 hour at RT, washed in phosphate buffered saline (PBS) and incubated with 0.1 to 0.3% H2O2 in PBS for 10 to 30 minutes. The tissue is again washed in PBS, then treated with an Avidin/Biotin blocking kit (Vector-cat #SP-2001), washed and incubated with BSA diluent for 1 hr at RT. The GFAP Antibody-W (1/30,000) is applied to the tissue overnight at RT. After washing in PBS the sections are incubated in Biotinylated GAR at 1/1000 (Vector labs-cat#BA1000) for 1hr at RT, washed and then incubated in Avidin/Biotin at 1/1000 (Vector labs-cat#PK6100) for 1hr at RT. After washing the signal is visualized with Diaminobenzidine (0.04%) in 0.1M Sodium Acetate with 0.06% H2O2 for six minutes (Nickel Chloride at 2.5% may also be added). Sections are then water washed (counterstained if desired), dehydrated and cover slipped.

GFAP Antibody-W also works with the protocol used to visualize the BRDU Antibody-W sold by Fluorochrome, LLC. However, 4% FA perfused tissue may be used instead of fresh frozen sections.



BrdU Antibody-W Use Guide and Protocol


General:
This antibody reacts against the synthetic thymidine analog BrdU, which when injected into an animal is incorporated during the S phase of the cell cycle into newly synthesized DNA strands of actively proliferating cells. The number of cells that survive can be measured by immunocytochemistry. As an outstanding tool to assess the proliferation state of a group of cells, it has become vital to drug discovery research, especially with cancer therapeutics and ascertaining the health of cells during ADME/Tox studies. BrdU Antibody-W is a polyclonal raised in rabbit against BrdU conjugated to bovine serum albumin (BSA). Our antibody has been used for immunocytochemistry in rat brain tissue. It will be sold with filter paper absorbed with BrdU for blocking. This allows the researcher to confirm the specific signal.

Characterization of the BrdU Antibody-W

美国fluorochrome471905,荧光金,Fluoro-Gold原装现货
A. BrdU positive cells in dentate gyrus (60X) of an adult rat previously injected with BrdU. Photo taken on confocal microscope
B. BrdU block in dentate gyrus

Specificity was ascertained by performing a blocking study. Adjacent sections from the hippocampus of an adult rat that had been previously injected with BrdU were incubated with the BrdU Antibody-W alone (A-60X) or with BrdU Antibody-W preincubated with BrdU (B-60X). Note labeling in A and the absence of labeling in B, indicating specificity of the antibody blocking.

Storage and Preparation:
The vial you will receive contains approximately 200μl at a dilution of 1/100. BrdU Antibody-W can be further diluted to at least 1/30,000 in BSA diluent (50mM KPBS, 0.4% Triton, 1% BSA, 1% NGS).

The antibody should be stored at -20C. Also included is filter paper absorbed with BrdU for blocking. 500μl of the diluted antibody can be added to the tube to achieve a 10 μM block. Incubate for at least 1hour at room temperature (RT) before applying to tissue. The antibody blocks specifically with BrdU and does not block with other peptides absorbed on filter paper.

Protocol:
Fresh frozen 10-30 micron sections are fixed in 4% paraformaldehyde (FA) for 1 hour at RT, washed in phosphate buffered saline (PBS), then treated with 50% Formamide/2X SSC (300 mM sodium chloride and 30 mM sodium citrate) at 65C for 2 hours. The sections are washed in 2X SSC, incubated in 2N HCL at 37C for 30 minutes, and then placed directly in 100mM Sodium Borate (pH 8.5) for 10 minutes at RT. The sections are washed in PBS and incubated with 0.1 to 0.3% H2O2 in PBS for 10 to 30 minutes. The tissue is again washed in PBS, then treated with an Avidin/Biotin blocking kit (Vector-cat #SP-2001), washed and incubated with BSA diluent for 1 hr at RT. The BrdU Antibody-W (1/30,000) is applied to the tissue overnight at RT. After washing in PBS the sections are incubated in Biotinylated GAR at 1/1000 (Vector labs-cat#BA1000) for 1hr at RT, washed and then incubated in Avidin/Biotin at 1/1000 (Vector labs-cat#PK6100) for 1hr at RT. After washing the signal is visualized with Diaminobenzidine (0.04%) in 0.1M Sodium Acetate with 0.06% H2O2 for six minutes (Nickel Chloride at 2.5% may also be added). Sections are then water washed (counterstained if desired), dehydrated and cover slipped.

Publications:
Garcia-Fuster et al, in preparation: Individual differences in adult hippocampal neurogensis following experimenter administration of cocaine.

Notice: The original and only true Fluoro-Gold (Fluorogold) is produced by Fluorochrome, LLC and marketed by Fluorochrome, LLC and Histo-Chem Inc.

Fluoro-Gold (Fluorogold) is an exclusive product of Fluorochrome, LLC. It has been sold by Fluorochrome and widely used since 1985. Other companies are marketing a product they claim is the same as or equivalent to Fluoro-Gold. In fact, the chemical structures of these compounds seem to be different from Fluoro-Gold. Certain physical properties of the compounds may be very different.

*CAUTION: Fluoro-Gold, Antibody to Fluoro-Gold and Fluoro-Ruby are for investigational use only in laboratory research animals or for tests in vitro. NOT FOR USE IN HUMANS. These drugs should be used only by persons regularly engaged in conducting neuroanatomical studies and tests in vitro or in animals used only for laboratory research

Antibody to Fluoro-Gold 报价
One vial 6243元
Two vial 10881元
Three vial 14242元
Each vial of the Antibody to Fluoro-Gold contains approximately 100μl of solution.