半岛bd体育手机客户端 说明
一般描述
Anti-ATM (Ab-3) (819-844), rabbit polyclonal, recognizes the ~350 kDa ATM protein in Daudi and HeLa cells. It is validated for Western blotting and immunoprecipitation.
Purified rabbit polyclonal antibody. Recognizes the ~350 kDa ATM protein.
Recognizes the ~350 kDa ATM protein in Daudi and HeLa cells.
免疫原
a synthetic peptide (CKSLASFIKKPFDRGEVESMEDDTNG) corresponding to amino acids 819-844 of human ATM
Human
包装
100 μg in Plastic ampoule
Please refer to vial label for lot-specific concentration.
应用
Immunoblotting (2 ?g/ml, see comments)
Immunoprecipitation (2 ?g)
警告
Toxicity: Standard Handling (A)
外形
In 50 mM sodium phosphate buffer, 0.2% gelatin.
分析说明
Negative Control
GM02052A cells
Positive Control
Daudi or HeLa cells
其他说明
Friedberg, E.C., et al. 1995.Amer. Soc. of Microbiolgy (meeting report), Wash. D.C.
Meyn, S.M. 1995.Cancer Res.55,5991.
Paules, R.S., et al. 1995.Cancer Res.55,1763.
Savitsky, K., et al. 1995.Science268,1749.
Savitsky, K., et al. 1995.Hum. Molec. Genet.4,2025.
Zakian, V., 1995.Cell82,685.
Beamish, H., et al. 1993.Rad. Res.138,130.
Kastan, M.B., et al. 1992.Cell71,587.
Painter, R.B. and Young, B.R. 1980.Proc. Natl. Acad. Sci. USA77,7315.
For immunoblotting use 100-150 ?g cell lysate on a 5% acrylamide gel (SDS/PAGE) and transferred to nitrocellulose using semi-dry transfer at 9V constant voltage for 2 h. Detection of antibody/antigen complexes is done using HRP conjugated goat anti-rabbit IgG at 25 ng/ml (Cat. No. DC03L).
1. Anti-ATM (Ab-3) Rabbit pAb immunoblots and immunoprecipitates a 350 kDa protein in lysates of normal and transformed cells or tumor lines derived from individuals homozygous wild type for the ATM gene; whereas in cells derived from patients with Ataxia-Telangiectasia and carrying homozygous inactivating mutations in ATM, no 350 kDa protein is detected.
2. Anti-ATM (Ab-3) Rabbit pAb may non-specifically detect smaller molecular weight proteins present in both ATM mutant and wild type cells. Careful titering of primary and secondary antibodies is recommended.
3. Immunoblotting of p350ATM requires loading 100-150 ?g cell lysate on low percentage acrylamide gels (5%) (SDS/PAGE) with electrophoresis performed until the 200 kDa molecular weight marker has migrated halfway through the gel. Semi-dry electrophoretic transfer is for 2 h at 9V constant voltage. Tank transfer is overnight at 40 V constant voltage.
4. To transfer the gel for blotting, lay a dry piece of Whatman 3MM Chromatography paper over the wet gel. Carefully peel the 3MM paper and gel off the glass plate and immerse, gel side up, in transfer buffer until 3MM paper is thoroughly wet. Remove bubbles by rolling a pipette across the surface of the gel.
5. To confirm detection of p350ATM, a cell line carrying a truncating mutation in ATM should be used as a negative control. Cell lines from A-T patients which show no detectable band at 350 kDa, can be obtained from the National Institute of General Medical Sciences Human Genetic Mutant Cell Repository at the Coriell Institute for Medical Research, 401 Haddon Avenue, Camden, NJ 08103.
6. For immunoprecipitations, prepare nuclear lysates as described. Immunoprecipitate p350ATM using 2 ?g of Anti-ATM (Ab-3) Rabbit pAb and Protein A-Agarose (Cat. No. IP06). Detection can be after metabolic labeling with35S methionine followed by autoradiography, or alternatively, immunoprecipitated proteins can be displayed on 5% SDS/PAGE, transferred to nitrocellulose and then blotted as above using Anti-ATM (Ab-3).
法律信息
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
半岛bd体育手机客户端 性质
生物来源 | rabbit |
质量水平 | 100 |
抗体形式 | purified antibody |
antibody product type | primary antibodies |
克隆 | polyclonal |
形式 | liquid |
包含 | ≤0.1% sodium azide as preservative |
species reactivity | human |
manufacturer/tradename | Calbiochem? |
储存条件 | do not freeze |
同位素/亚型 | IgG |
运输 | wet ice |
储存温度 | 2-8℃ |
安全信息
储存分类代码 | 12 - Non Combustible Liquids |
WGK | nwg |
闪点(F) | Not applicable |
闪点(C) | Not applicable |
Sigma-Aldrich