Pro-Survival Bcl-2 Family Antibody Sampler Kit II

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Pro-Survival Bcl-2 Family Antibody Sampler Kit II

品牌： CST

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产品信息

荧光素标记

Unconjugated

抗原名称

Pro-Survival Bcl-2 Family

简单描述

The Pro-Survival Bcl-2 Family Antibody Sampler Kit II provides an economical means to examine several members of the Bcl-2 family. The kit contains enough primary antibody to perform two western blot experiments.

研究领域

癌症,细胞生物学,纤维化,神经科学,

背景

The Bcl-2 family consists of a number of evolutionarily conserved proteins containing Bcl-2 homology domains (BH) that regulate apoptosis through control of mitochondrial membrane permeability and release of cytochrome c (1-3). Four BH domains have been identified (BH1-4) that mediate protein interactions. The family can be separated into three groups based upon function and sequence homology: pro-survival members include Bcl-2, Bcl-xL, Mcl-1, A1 and Bcl-w; pro-apoptotic proteins include Bax, Bak and Bok; and "BH3 only" proteins Bad, Bik, Bid, Puma, Bim, Bmf, Noxa and Hrk. Interactions between death-promoting and death-suppressing Bcl-2 family members has led to a rheostat model in which the ratio of pro-apoptotic and anti-apoptotic proteins controls cell fate (4). Thus, pro-survival members exert their behavior by binding to and antagonizing death-promoting members. In general, the "BH3-only members" can bind to and antagonize the pro-survival proteins leading to increased apoptosis (5). While some redundancy of this system likely exists, tissue specificity, transcriptional and post-translational regulation of many of these family members can account for distinct physiological roles.Several phosphorylation sites have been identified within Bcl-2 including Thr56, Ser70, Thr74 and Ser87 (6). These phosphorylation sites may be targets of the ASK1/MKK7/JNK1 pathway, and phosphorylation of Bcl-2 may be a marker for mitotic events (7,8). Mutation of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity during glucocorticoid-induced apoptosis of T lymphocytes (9). Interleukin 3 and JNK-induced Bcl-2 phosphorylation at Ser70 may be required for its enhanced antiapoptotic functions (10).Mcl-1 is phosphorylated in response to treatment with phorbol ester, microtubule-damaging agents, oxidative stress, and cytokine withdrawal (11-14). Phosphorylation at Thr163, the conserved MAP kinase/ERK site located within the PEST region, slows Mcl-1 protein turnover (13) but may prime the GSK-3 mediated phosphorylation at Ser159 that leads to Mcl-1 destabilization (14). 1.Cory, S. et al. (2003) Oncogene 22, 8590-607. 2.Antonsson, B. and Martinou, J.C. (2000) Exp Cell Res 256, 50-7. 3.Sharpe, J.C. et al. (2004) Biochim Biophys Acta 1644, 107-13. 4.Korsmeyer, S.J. et al. (1993) Semin Cancer Biol 4, 327-32. 5.Bouillet, P. and Strasser, A. (2002) J Cell Sci 115, 1567-74. 6.Maundrell, K. et al. (1997) J Biol Chem 272, 25238-42. 7.Yamamoto, K. et al. (1999) Mol Cell Biol 19, 8469-78. 8.Ling, Y.H. et al. (1998) J Biol Chem 273, 18984-91. 9.Huang, S.T. and Cidlowski, J.A. (2002) FASEB J 16, 825-32. 10.Deng, X. et al. (2001) J Biol Chem 276, 23681-8. 11.Domina, A.M. et al. (2000) J Biol Chem 275, 21688-94. 12.Inoshita, S. et al. (2002) J Biol Chem 277, 43730-4. 13.Domina, A.M. et al. (2004) Oncogene 23, 5301-15. 14.Maurer, U. et al. (2006) Mol Cell 21, 749-60.

研究领域

癌症,细胞生物学,纤维化,神经科学,

翻译后修饰

phosphate

数据库链接

Entrez-Gene ID

4170,598,17210,596,599,597

UniProt ID

Q07820,Q07817,P97287,P10415,Q92843,Q16548

参考图片

Simple Western™ analysis of lysates (1 mg/mL) from MCF-7 cells using Mcl-1 (D2W9E) Rabbit mAb #94296. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human A1/Bfl-1 protein (hA1/Bfl-1; +), using A1/Bfl-1 (D1A1C) Rabbit mAb.

Western blot analysis of extracts from KARPAS-299 and Raji cell lines, untreated (-) or treated with TPA #4174 (200 nM, 30 min; +), using Phospho-Mcl-1 (Thr163) (D5M9D) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). KARPAS cell Line source: Dr Abraham Karpas at the University of Cambridge.

Western blot analysis of extracts from A673 and C2C12 cell lines, using Bcl-w (31H4) Rabbit mAb.

Western blot analysis of extracts from Jurkat and HeLa (human), COS (monkey), NIH/3T3 and L929 (mouse), and PC12 and C6 (rat) cells, using Bcl-xL (54H6) Rabbit mAb.

Flow cytometric analysis of THP-1 cells (blue, moderate expression) and K-562 cells (green, high expression) using Bcl-xL (54H6) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Western blot analysis of extracts from Jurkat cells, untreated or treated with paclitaxel (1 μM, overnight) and with or without λ phosphatase, using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb (upper) or Bcl-2 #2876 (lower).

Western blot analysis of control HeLa cells (lane 1) or Bcl-2 knockout HeLa cells (lane 2) using Bcl-2 (D55G8) Rabbit mAb #4223 (upper), or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Bcl-2 knockout HeLa cells confirms the specificity of the antibody for Bcl-2.

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Western blot analysis of extracts from 293T cells, mock transfected or transfected with a construct expressing full-length human Mcl-1 (hMcl-1; +) or mouse Mcl-1 (mMcl-1; +), using Mcl-1 (D2W9E) Rabbit mAb.

Western blot analysis of extracts from U-937 and UACC-62 cells using A1/Bfl-1 (D1A1C) Rabbit mAb.

Immunoprecipitation of phospho-Mcl-1 (Thr163) from KARPAS-299 cells treated with TPA #4174 (200 nM, 30 min) using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-Mcl-1 (Thr163) (D5M9D) Rabbit mAb (lane 3). Lane 1 represents 10% input. Western blot was perform using Phospho-Mcl-1 (Thr163) (D5M9D) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody to avoid cross reactivity with IgG heavy and light chains. KARPAS cell Line source: Dr Abraham Karpas at the University of Cambridge.

Western blot analysis of recombinant Bcl-w (amino acids 1-172) using Bcl-w (31H4) Rabbit mAb.

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® BcL-xL siRNA I (+) or SignalSilence® Bcl-xL siRNA II #6363 (+), using Bcl-xL (54H6) Rabbit mAb #2764 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Bcl-xL (54H6) Rabbit mAb confirms silencing of Bcl-xL expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Western blot analysis of extracts from various cell lines using Bcl-2 (D55G8) Rabbit mAb.

Immunoprecipitation of A1/Bfl-1 from Mutz-3 cell extracts. Lane 1 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lane 2 is 10% input, and lane 3 is A1/Bfl-1 (D1A1C) Rabbit mAb #14093. Western blot analysis was performed using A1/Bfl-1 (D1A1C) Rabbit mAb #14093. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.

Immunoprecipitation of Bcl-2 from ACHN extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Bcl-2 (D55G8) Rabbit mAb. Western blot analysis was performed using Bcl-2 (124) Mouse mAb #15071. Anti-mouse IgG, HRP-linked Antibody #7076 was used as a secondary antibody.

Western blot analysis of extracts from various cell lines using Mcl-1 (D2W9E) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Immunoprecipitation of Bcl-xL from Jurkat cell extracts, using Bcl-xL (54H6) Rabbit mAb. Lane 1 is the lysate control, lane 2 is antibody alone and lane 3 is antibody plus lysate.

Simple Western™ analysis of lysates (1.0 mg/mL) from HeLa cells using Bcl-xL (54H6) Rabbit mAb #2764. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Immunoprecipitation of Mcl-1 from MCF7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Mcl-1 (D2W9E) Rabbit mAb. Western blot analysis was performed using Mcl-1 (D2W9E) Rabbit mAb. A conformation-specific secondary antibody was used to avoid cross-reactivity with IgG.

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Bcl-xL (54H6) Rabbit mAb.

Flow cytometric analysis of Jurkat cells using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087 to measure DNA content. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Confocal immunofluorescent analysis of MCF7 (left), L-929 (center), and SK-OV-3 (right) cells using Mcl-1 (D2W9E) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Bcl-xL (54H6) Rabbit mAb in the presence of control peptide (left) or Bcl-xL Blocking Peptide #1225 (right).

Flow cytometric analysis of L-929 cells using Mcl-1 (D2W9E) Rabbit mAb (solid line) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, showing cytoplasmic localization, using Bcl-xL (54H6) Rabbit mAb.

Flow cytometric analysis of SK-OV-3 cells (low expression; blue) and MCF7 cells (high expression; green) using Mcl-1 (D2W9E) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Confocal immunofluorescent analysis of HeLa cells using Bcl-xL (54H6) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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货号：

17229T

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