- Description
- Additional
Description
Details
Description:Mouse monoclonal antibody to human Raf-1
Purification:Protein G affinity purified
Product Type:Primary antibody
Target Protein:Human Raf-1
Immunogen:Purified recombinant Raf-1 (mw:75kDa)
Fusion Myeloma:Sp2/0-Ag14
Specificity:Reactive to Raf-1 N-terminal (1-240 a. a.)
Species Reacitvity:Human, others not tested
Host / Isotype: Mouse, IgG1 Kappa
Formulation:Lyophilized from a solution in 0.01M PBS pH7.2
Reconstitution:Double distilled water is recommended to adjust the final concentration to 1.00mg/mL.
Storage:Store at -20oC
Research Area:Cell differentiation, oncology
Background:
Raf-1 is a serine/threonine protein kinase in the mitogen-activated protein kinase (MAPK) pathway. When extracellular signal binds to the G-protein coupled membrane receptor, Ras binds to GTP and phosphorylates Raf. The activated Raf phosphorylates MAPK kinase, which consecutively, activates MAPK. MAPK triggers the transcription of a wide spectrum of genes involved with cell growth and differentiation by activating transcription factors such as C-myc andCREB. MAPK also regulates translation by phosphorylating RSK, a ribosomal protein 6S kinase.
Gene for Raf-1 is located on chromosome 3. The mutations on Raf-1 may relate to some rare but severe diseases such as Noonan syndrome and Leopard syndrome.
Application:
Western Blot:The antibody, when used at 1μg/mL, will allow visualization of 0.2μg/lane of recombinant human Raf-1 (75 kDa).
References:
If research is published using this product, please inform Anogen in order to cite the reference on this datasheet. Anogen will provide one unit of product in the same category as gratitude.
Additional
Additional Information
| Product Specificity | mAb anti-Human Raf-1, 410 |
|---|---|
| Application | WB |
| Size | 0.5 mg |
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尤其是获得SCI文献引用的产品，
绝对ok的
用Mabteck公司的ALP标记抗体吧，配合Millipore公司的Multiscreen-IP96孔板，所有的试剂都可以减半，照样可以出非常好的结果，4*96孔的试剂盒我一直做8*96孔，但BCIP/NBT显色液没试过减半，因为它很便宜。记得当年第一次做实验就出了很好的结果，经过不断摸索，做出的蓝紫色斑点确实非常漂亮，几乎没有非特异斑点，嗯，ELISPOT是我最拿手的实验之一。
关于PBMC:
拿到血液标本后，首先我要做的是尽快分离PBMC（一般2h内），国产的分离液里天津灏洋的比较好，然后用冻存液冻存PBMC。PBMC的冻存过程比一般的细胞株要更为严格，最好用冻存器。ELISPOT试验的第一天，在包被抗体的同时，将细胞复苏，完全培养液培养过夜，第二天收集细胞并计数即可。
请问一般的实验室可以进行ELIspot吗？（我可以买到现成的试剂盒，资金这个应该没有问题。）ELIspot的难点在哪里？多谢大虾们的指点！这个对我很重要，多谢！！！
我想用酶联免疫斑点（ELISPOT）技术测定NK细胞活性，但不知道哪里能买到试剂盒，特向大家求助，谢谢。
