Fluorochrome荧光金抗体,AntibodytoFluoro-Gold,Ki-67Antibody-W,GFAPAntibody-W,BrdUAntibody-W

Our Antibody to Fluoro-Goldis by far the most effective antibody to Fluoro-Gold available.Since Fluorochrome first introduced it in 1998, it has a achieved aproven record of accomplishment and has become the standard in theindustry.

The Ki-67 Antibody-W, GFAPAntibody-W, and BrdU Antibody-W are excellent antibodies that arerecent introductions. Included with these antibodies is filterpaper absorbed with a specific blocking peptide. We feel that ablocking agent should be utilized in all investigations using thesetypes of antibodies. Only with a blocking agent can the researcherverify and have confidence that the observed signal is the intendedsignal. At no additional costs, we have included the blockingagent, which makes this verification easy and accurate for theresearcher to implement.
As an introductory offer, we are discounting the purchase price forthe Ki-67 Antibody-W, GFAP Antibody-W, and BrdU Antibody-W by20%.

Antibody to Fluoro-Gold in a small vial. It is a polyclonalantibody raised in rabbit. The vial will contain approximately100 l of theantibody solution. It should be immediately frozen and stored in acold (-20 degrees C), dark, dry environment. The freezer section,without the frost free feature, of a good commercial refrigeratorshould suffice. If properly and continuously frozen, the antibodysolution can be stored up to one year.
The antibody solution should remain frozen until ready for use. The100 l aliquot ofthe Antibody is at a dilution of 1/100. The Antibody can be furtherdiluted 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 workingtiter. This means you can dilute the solution you receive by 500 to1,000 times. This should treat 300 to 1000 sections if you use theVector elite kit. After preparation, the antibody solution can bestored for up to seven days in a cool (4 degrees C), dark and dryenvironment. The refrigerator section of a good commercialrefrigerator should suffice. We do not recommend that the solutionbe used beyond seven days after preparation. Do not refreeze theantibody solution.
Use of theAntibody
Fluoro-Gold can be injected using several different methods,including pressure, iontophoretic and other applications developedby a variety of researchers. See Schmued and Fallon,Fluoro-Gold: \"A fluorescent retrograde axonal tracer with numerousunique properties,\" Brain Research, 377 (1986) 147-154 as wellas Pieribone and Aston-Jones, \"The Iontophoretic Application ofFluoro-Gold for the study of afferents to deep brain nuclei,\" BrainResearch, 475 (1988) 259-271. Many researchers have developedtheir own modified procedures. Use of the antibody should not bedependent upon the methodology used to employ Fluoro-Gold.
After the Fluoro-Gold has been injected, floating sections (we usedthirty um sections from a rat perfused with 4% formaldehyde) areincubated with the Fluoro-Gold Antibody solution overnight at 4degrees C. Sections are washed, then incubated in Biotinylated GAR(Vector Labs) at 1/1000 for 1 hour at room temperature and washedagain. 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 thenwashed, mounted, dried, dehydrated and cover slipped.
It has been our experience that if stored and prepared in themanner set out above, each vial of the antibody should treat 300 to1000 thirty um sections of the albino rat brain (or similar sizedanimals) if you use the Vector elite kit. However, you shouldexperiment with the concentration and procedures to determine whichbest fits your circumstance.

Ki-67 Antibody-W UseGuide and Protocol
General:
Ki-67 Antibody-W reacts againstthe Ki-67 protein. The Ki-67 protein is anintrinsic marker of ongoing cell proliferation and is presentduring all active phases of the cell cycle (G1, S, G2 and mitosis)and is absent from resting cells (G0). This meansit is an excellent marker for determining the growth fraction of agiven cell population. It is used extensively indiagnostic, research and drug discovery applications. Ki-67Antibody-W is polyclonal and was raised in rabbit to the humanpeptide sequence TPKEKAQALEDLAGFKELFQT that was coupled tothyroglobulin by glutaraldehyde before injection into the rabbit.Our antibody has been used for immunocytochemistry in rat and humanbrain tissue. It will be sold with filter paperabsorbed with the specific peptide for blocking. This allows theresearcher to confirm the specific signal.
Characterization of the Ki-67Antibody-W

Specificity was ascertained by performing a blocking study.Adjacent sections from the hippocampus of a 7 day old rat wereincubated with the Ki-67 Antibody-W alone (A-10X) and (B-60X) orwith 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 andPreparation:
The vial you will receive contains approximately 200 l of solutionat a dilution of 1/100. The Ki-67Antibody-W can be further dilutedto 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 filterpaper absorbed with the specific peptide for blocking. 500 l of thediluted antibody can be added to the tube to achieve a 10 M block.Incubate for at least 1 hour at room temperature (RT) beforeapplying to tissue. The antibody blocks specifically with thispeptide and does not block with other peptides absorbed on filterpaper.
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. Thesections are allowed to cool for 20 minutes in the sodium citratesolution, washed in PBS and incubated with 0.1 to 0.3% H2O2 in PBSfor 10 to 30 minutes. The tissue is again washed in PBS, thentreated with an Avidin/Biotin blocking kit (Vector-cat #SP-2001),washed and incubated with BSA diluent for 1 hour at RT. The Ki67antibody (1/20,000) is applied to the tissue overnight at RT. Afterwashing in PBS the sections are incubated in Biotinylated GAR at1/1000 (Vector labs-cat#BA1000) for 1hr at RT, washed and thenincubated in Avidin/Biotin at 1/1000 (Vector labs-cat#PK6100) for1hr at RT. After washing the signal is visualized withDiaminobenzidine (0.04%) in 0.1M Sodium Acetate with 0.06% H2O2 forsix minutes (Nickel Chloride at 2.5% may also be added). Sectionsare then water washed (counterstained if desired), dehydrated andcover slipped.
Ki-67 Antibody-W also works with the protocol used to visualize theBRDU Antibody-W sold by Fluorochrome, LLC. However, 4% FA perfusedtissue 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 followingexperimenter administration of cocaine.
GFAP Antibody-W reacts againstthe Glial Fibrillary Acidic Protein (GFAP), which is a class-IIIintermediate filament and the main constitute of intermediatefilaments in astrocytes and serves as a cell specific marker formature glial cells. It is useful as a marker ofneural stem cells and astrocytic cells, including many types ofbrain tumor derived from astrocytic cells, which expressGFAP. GFAP Antibody-W is polyclonal and wasraised in rabbit to the peptide sequence NAGFKETRASERAE (human, ratand mouse) coupled to thyroglobulin by glutaraldehyde beforeinjection into the rabbit. Our antibody has been used forimmunocytochemistry in rat and human braintissue. It will also be sold with filter paperabsorbed with the specific peptide for blocking.This allows the researcher to confirm the specificsignal.
Characterization of the GFAPAntibody-W

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

Storage andPreparation:
The vial you will receive contains approximately200 l at a dilutionof 1/100. The antibody can be further diluted to at least 1/30,000in BSA diluent (50mM KPBS, 0.4% Triton, 1% BSA, 1% NGS).
The antibody should be stored at -20C. Also included is filterpaper absorbed with the specific peptide for blocking. 500 l of thediluted antibody can be added to the tube to achieve a 10 M block.Incubate for at least 1 hour at room temperature (RT) beforeapplying to tissue. The antibody blocks specifically with thispeptide and does not block with other peptides absorbed on filterpaper.
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 anAvidin/Biotin blocking kit (Vector-cat #SP-2001), washed andincubated with BSA diluent for 1 hr at RT. The GFAP Antibody-W(1/30,000) is applied to the tissue overnight at RT. After washingin PBS the sections are incubated in Biotinylated GAR at 1/1000(Vector labs-cat#BA1000) for 1hr at RT, washed and then incubatedin 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 thenwater washed (counterstained if desired), dehydrated and coverslipped.
GFAP Antibody-W also works with the protocol used to visualize theBRDU Antibody-W sold by Fluorochrome, LLC. However, 4% FA perfusedtissue may be used instead of fresh frozensections.

This antibody reacts against thesynthetic thymidine analog BrdU, which when injected into an animalis incorporated during the S phase of the cell cycle into newlysynthesized DNA strands of actively proliferatingcells. The number of cells that survive can bemeasured by immunocytochemistry. As anoutstanding tool to assess the proliferation state of a group ofcells, it has become vital to drug discovery research, especiallywith cancer therapeutics and ascertaining the health of cellsduring ADME/Tox studies. BrdU Antibody-W is apolyclonal raised in rabbit against BrdU conjugated to bovine serumalbumin (BSA). Our antibody has been used for immunocytochemistryin rat brain tissue. It will be sold with filterpaper absorbed with BrdU for blocking. Thisallows the researcher to confirm the specific signal.
Characterization of the BrdUAntibody-W

positive cells in dentate gyrus (60X) of an adult rat previouslyinjected with BrdU. Photo taken on confocal microscope
Specificity was ascertained by performing a blocking study.Adjacent sections from the hippocampus of an adult rat that hadbeen previously injected with BrdU were incubated with the BrdUAntibody-W alone (A-60X) or with BrdU Antibody-W preincubated withBrdU (B-60X). Note labeling in A and the absence of labeling in B,indicating specificity of the antibody blocking.

Storage andPreparation:
The vial you will receive contains approximately 200 l at adilution of 1/100. BrdU Antibody-W can be further diluted to atleast 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 filterpaper absorbed with BrdU for blocking. 500 l of the dilutedantibody can be added to the tube to achieve a 10 M block.Incubate for at least 1hour at room temperature (RT) beforeapplying to tissue. The antibody blocks specifically with BrdU anddoes 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 (300mMsodium chloride and 30mM sodium citrate) at 65Cfor 2 hours. The sections are washed in 2X SSC, incubated in 2N HCLat 37C for 30 minutes, and then placed directly in 100mM SodiumBorate (pH 8.5) for 10 minutes at RT. The sections are washed inPBS and incubated with 0.1 to 0.3% H2O2 in PBS for 10 to 30minutes. The tissue is again washed in PBS, then treated with anAvidin/Biotin blocking kit (Vector-cat #SP-2001), washed andincubated with BSA diluent for 1 hr at RT. The BrdU Antibody-W(1/30,000) is applied to the tissue overnight at RT. After washingin PBS the sections are incubated in Biotinylated GAR at 1/1000(Vector labs-cat#BA1000) for 1hr at RT, washed and then incubatedin 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 thenwater washed (counterstained if desired), dehydrated and coverslipped.
Publications:
Garcia-Fuster et al, in preparation: Individual differences inadult hippocampal neurogensis following experimenter administrationof cocaine.

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

Fluoro-Gold (Fluorogold)is an exclusive product of Fluorochrome, LLC. It has been sold byFluorochrome and widely used since 1985. Other companies aremarketing a product they claim is the same as orequivalent to Fluoro-Gold. In fact, the chemical structures ofthese compounds seem to be different from Fluoro-Gold. Certainphysical properties of the compounds may be verydifferent.
*CAUTION: Fluoro-Gold, Antibody to Fluoro-Gold andFluoro-Ruby are for investigational use only in laboratory researchanimals or for tests in vitro. NOT FOR USE INHUMANS. These drugs should be used only by persons regularlyengaged in conducting neuroanatomical studies and tests in vitro orin animals used only for laboratory research.