| 货号 | 12854T |
| 描述 | The SWI/SNF Complex Antibody Sampler Kit provides an economical means of detecting total protein from the SWI/SNF family members including ARID1A/BAF250A, Brg1, BRM, SMARCC1/BAF155, SMARCC2/BAF170 and SNF5. The kit contains enough primary antibody to perform four western blots per primary antibody. |
| 目标/特异性 | Each antibody in this kit recognizes endogenous levels of total protein for the specified target and does not cross-react with other family members. ARID1A/BAF250A (D2A8U) Rabbit mAb also cross-reacts with proteins of unknown origin at 65 kDa. |
| 供应商 | CST |
| 背景 | ATP-dependent chromatin remodeling complexes play an essential role in the regulation of various nuclear processes, such as gene expression, DNA replication, and repair (1,2). The SWI/SNF chromatin remodeling complex consists of more than 10 subunits with a single molecule of the ATPase catalytic subunit BRM or BRG1, but not both. The activities of these two subunits drive the disruption of histone-DNA contacts that lead to changes in accessibility of crucial regulatory elements within chromatin (2-5). The BRM/BRG1 containing SWI/SNF complexes are recruited to target promoters by transcription factors, such as nuclear receptors, p53, RB, and BRCA1 to regulate gene activation, cell growth, the cell cycle, and differentiation processes (1,6-9). BRM and BRG1 are also considered to be tumor suppressors and their expression levels are severely reduced in several cancer cell lines (10-13). SMARCC1/BAF155, SMARCC2/BAF170, and SNF5 are members of the core subunits of the SWI/SNF complex, which is necessary for efficient nucleosome remodeling by BRG1 in vitro (14). ARID1A/BAF250A is one of the accessory subunits of the SWI/SNF complex (15). SMARCC1, SNF5, and ARID1A are an essential part of the mouse embryonic stem cell specific SWI/SNF complex (esBAF). SMARCC1 is necessary for early embryogenesis, especially proper brain and visceral endoderm development (16-18). SNF5 is necessary for early embryogenesis and hepatocyte differentiation (19,20). ARID1A is critical for ES cell pluripotency and differentiation into mesoderm-derived cardiomyocytes and adipocytes (15). While SMARCC2 has been shown to be part of the SWI/SNF complex in non-pluripotent cells, it is absent in pluripotent embryonic stem (ES) cells. Expression of SMARCC2 has been shown to be up-regulated in neurons/neuronal progenitors upon differentiation of mouse ES cells with retinoic acid, and exogenous expression of SMARCC2 leads to loss of stem cell pluripotency and self renewal (21). |
| 存放说明 | -20C |
| 参考文献 | 1 . Ho, L. and Crabtree, G.R. (2010) Nature 463, 474-84. 2 . Becker, P.B. and Hörz, W. (2002) Annu Rev Biochem 71, 247-73. 3 . Eberharter, A. and Becker, P.B. (2004) J Cell Sci 117, 3707-11. 4 . Bowman, G.D. (2010) Curr Opin Struct Biol 20, 73-81. 5 . Gangaraju, V.K. and Bartholomew, B. (2007) Mutat Res 618, 3-17. 6 . Lessard, J.A. and Crabtree, G.R. (2010) Annu Rev Cell Dev Biol 26, 503-32. 7 . Morettini, S. et al. (2008) Front Biosci 13, 5522-32. 8 . Wolf, I.M. et al. (2008) J Cell Biochem 104, 1580-6. 9 . Simone, C. (2006) J Cell Physiol 207, 309-14. 10 . Yamamichi, N. et al. (2005) Oncogene 24, 5471-81. 11 . Phelan, M.L. et al. (1999) Mol Cell 3, 247-53. 12 . Reisman, D.N. et al. (2002) Oncogene 21, 1196-207. 13 . Ho, L. et al. (2009) Proc Natl Acad Sci U S A 106, 5181-6. 14 . Shen, H. et al. (2008) Cancer Res 68, 10154-62. 15 . Weissman, B. and Knudsen, K.E. (2009) Cancer Res 69, 8223-30. 16 . Gao, X. et al. (2008) Proc Natl Acad Sci U S A 105, 6656-61. 17 . Han, D. et al. (2008) Dev Biol 315, 136-46. 18 . Kim, J.K. et al. (2001) Mol Cell Biol 21, 7787-95. 19 . Schaniel, C. et al. (2009) Stem Cells 27, 2979-91. 20 . Klochendler-Yeivin, A. et al. (2000) EMBO Rep 1, 500-6. 21 . Gresh, L. et al. (2005) EMBO J 24, 3313-24. |
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 of various cell lines using Brg1 (A52) Antibody. | |
Confocal immunofluorescent analysis of 293 cells using Brg1 (A52) Antibody (green). Actin filaments have been labeled using DY-554 phalloidin (red). | |
Western blot analysis of extracts from various cell lines using SNF5 (D9C2) Rabbit mAb. | |
Western blot analysis of extracts from various cell lines using SMARCC1/BAF155 (D7F8S) Rabbit mAb. | |
Immunoprecipitation of SMARCC1/BAF155 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or SMARCC1/BAF155 (D7F8S) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using SMARCC1/BAF155 (D7F8S) Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells, grown in phenol red-free medium and 5% charcoal-stripped FBS for 4 d followed by treatment with β-estradiol (10 nM, 45 min), and either 5 μl of SMARCC1/BAF155 (D7F8S) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Confocal immunofluorescent analysis of HeLa (positive, left) and NCCIT (negative, right) cells using BRM (D9E8B) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d followed by treatment with β-estradiol (10 nM, 45 min) and either 5 μl of BRM (D9E8B) XP® Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Western blot analysis of extracts from various cell lines using BRM (D9E8B) XP®Rabbit mAb (upper) or Brg1 (A52) Antibody #3508 (lower). | |
Western blot analysis of extracts from various cell lines using ARID1A/BAF250A (D2A8U) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded cell pellets, COS-7 (left) or T-47D (right), using ARID1A/BAF250A (D2A8U) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human lung adenosquamous carcinoma using ARID1A/BAF250A (D2A8U) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). | |
Western blot analysis of T-47D and Jurkat cell extracts using ARID1A/BAF250A (D2A8U) Rabbit mAb (upper) or ß-Actin (D6A8) Rabbit mAb #8457 (lower). Additional ARID1A/BAF250A degradation products may be detected in some cell extracts between 135kDa-250kDa, which are absent in the ARID1A/BAF250A negative T-47D cell line. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells, grown in phenol red-free medium and 5% charcoal-stripped FBS for 4 d followed by treatment with β-estradiol (10 nM, 45 min), and either 10 μl of SMARCC2/BAF170 (D8O9V) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. |
