Product Specifications:
Item # 1301-F: Fluorescene Rabbit Anti-gp120 HIV-1 IIIB pAb IgG
Mass/vial: Approx. 50ug IgG
Vol/vial: 100ul
Diluent: 10mM Tris, pH 8.0
Stabilizer: None
Preservative: None
Storage: -75°C
F/P Ratio: 2 - 4
Physical State: Frozen Liquid
Stability: At least 12 months at -75°C
Minimum Dilution: 1 : 50
Applications: Immunohistocytology, Immunostaining/sorting, Dot Blots, FACS.
Description: Fluorescene conjugated Rabbit anti-gp120 (HIV-1 IIIB) pAb IgG.
Purification: Highly purified mAb IgG (>95% pure) was used for FITC conjugation and the conjugate purified by gel exclusion chromatography.
Specificity: This fluorescene conjugated pAb IgG binds to native and recombinant HIV-1 gp120 in Dot Blot assays and stains cell surfaces in direct immunofluorescence assays.
Biological Activity: None
Application and Instruction for use:
Recommended dilutions for use are approximate values. A dose dependent response assay should be performed to determine the optimal concentration for use in specific applications. Direct Immunofluorescent staining of cells may be performed in 1 : 50 dilution range. Dot Blot assays with purified gp120 may be performed at a minimum dilution of 1 : 100.
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1、如果是提取的总蛋白，然后做WB，用β-actin或者GAPDH做内参肯定是没有问题的，这是公认的东西。
2、如果用膜蛋白提取试剂盒提取蛋白，再用β-actin作为内参似乎不妥，因为理论上来讲β-actin在膜上是不表达的。WB能做出β-actin来是因为膜蛋白提取时把胞质蛋白也提出来了。然而，如果我们试验目的是用药物处理细胞，比较处理前后某种膜蛋白的表达情况，此时用膜蛋白提取试剂盒提取膜蛋白后再用β-actin做内参似乎就不妥了，因为你根本不知道药物处理前后混杂了多少的胞质蛋白进来。如果没有胞质蛋白混进去的话，β-actin就是检测不到的。
转膜后就要区分与胶接触的一面与另一面了。话说楼主好奇的话可以跑蛋白的试试看哈。比如目的蛋白的胶用光滑面贴着胶，内参蛋白用粗糙面贴着胶一起转膜试试转膜结果看看呢。
