艾美捷GFAP抗体应用类型及保存建议

GFAP is a marker for mature glial cells. This antibody is polyclonal and was raised in rabbit to the peptide sequence – NAGFKETRASERAE (human, rat and mouse) – that was coupled to thyroglobulin by glutaraldehyde before being injected into the rabbit. The antibody has been used for immunocytochemistry in rat and human brain tissue. The vial contains about 200ul 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. 500ul of the diluted antibody can be added to the tube to achieve a 10 uM block. Incubate for at least 1hr 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.

GFAP是一种3型中间丝作为中枢神经系统发育期间的细胞特异性标记以区分星形胶质细胞与其他胶质细胞。GFAP抗体已经被广泛证实仅识别星形胶质细胞(astrocyte,也称astroglia),而不识别神经细胞、成纤维细胞、少突胶质细胞(oligodendrocyte)以及肿瘤细胞等其它类型的细胞。

艾美捷GFAP抗体是多克隆的，在兔体内培养至肽序列NAGFKETRASERAE（人、大鼠和小鼠），通过戊二醛，然后注射到兔子体内。该抗体已用于大鼠和人脑组织的免疫细胞化学。

小瓶中含有约200ul，稀释度为抗体可在BSA稀释剂（50mM KPBS、0.4%Triton、1%BSA、1%NGS）中进一步稀释至至少1/30000。抗体应储存在-20℃。还包括滤纸用于阻断的特异性肽。可将500ul稀释的抗体加入试管中uM块。在室温（RT）下孵育至少1小时，然后施用于组织。抗体与该肽特异性阻断，而不与滤纸上吸收的其他肽阻断。

GFAP抗体（FC50001）基本参数：

中文名称：GFAP抗体

英文名字：GFAP Antibody -W

cat#：FC50001

规格：200ul

应用类型：IHC

免疫原：这种抗体是多克隆抗体，在兔体内产生的肽序列是NAGFKETRASERAE（人、大鼠和小鼠），在注射到兔体内之前，用戊二醛将其与甲状腺球蛋白偶联。

来源宿主：兔

反应性：人，大鼠

保存建议：小瓶中含有约200微升1/100稀释液。 抗体可在BSA稀释液（50 mM KPBS、0.4%Triton、1%BSA、1%NGS）中进一步稀释至至少1/20，000。 抗体应在-20 ℃下储存。 还包括吸附有用于封闭的特异性肽的滤纸。 可向管中加入500 μ l稀释的抗体以获得10 μ M阻断。 应用于组织前，在室温（RT）下孵育至少1小时。 抗体特异性地与该肽阻断，而不与吸附在滤纸上的其他肽阻断。

GFAP抗体（FC50001）基本操作：

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. TheGFAP antibody (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抗体（FC50001）也适用于BRDU抗体可视化方案。可以使用4%FA灌注组织代替新鲜冷冻切片。