| 货号 | 4356T |
| 反应种属 | Human,Mouse,Rat,Monkey, |
| 来源宿主 | Rabbit |
| 应用 | WB, IP , IHC-P , F , IF-IC , ChIP |
| 目标/特异性 | This antibody detects endogenous levels of total HSF1 protein. |
| 使用方法 | WB(1:1000) IP (1:50) IHC-P (1:250) F (1:50) IF-IC (1:500) ChIP (1:50) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | All organisms respond to increased temperatures and other environmental stresses by rapidly inducing the expression of highly conserved heat shock proteins (HSPs) that serve as molecular chaperones to refold denatured proteins and promote the degradation of damaged proteins. Heat shock gene transcription is regulated by a family of heat shock factors (HSFs), transcriptional activators that bind to heat shock response elements (HSEs) located upstream of all heat shock genes (1). HSEs are highly conserved among organisms and contain multiple adjacent and inverse iterations of the pentanucleotide motif 5-nGAAn-3. HSFs are less conserved and share only 40% sequence identity. Vertebrate cells contain four HSF proteins: HSF1, 2 and 4 are ubiquitous, while HSF3 has only been characterized in avian species. HSF1 induces heat shock gene transcription in response to heat, heavy metals, and oxidative agents, while HSF2 is involved in spermatogenesis and erythroid cell development. HSF3 and HSF4 show overlapping functions with HSF1 and HSF2. The inactive form of HSF1 exists as a monomer and localizes to both the cytoplasm and nucleus, but does not bind DNA (1,2). In response to stress, HSF1 becomes phosphorylated, forms homotrimers, binds DNA and activates heat shock gene transcription (1,2). HSF1 activity is positively regulated by phosphorylation of serine 419 by PLK1, which enhances nuclear translocation, and phosphorylation of serine 230 by CaMKII, which enhances transactivation (3,4). Alternatively, HSF1 activity is repressed by phosphorylation of serines 303 and 307 by GSK3 and ERK1, respectively, which leads to binding of 14-3-3 protein and sequestration of HSF1 in the cytoplasm (5,6). In addition, during attenuation from the heat shock response, HSF1 is repressed by direct binding of Hsp70, HSP40/Hdj-1, and HSF binding protein 1 (HSBP1) (7). |
| 存放说明 | -20C |
| 计算分子量 | 82 |
| 参考文献 | 1 . Morimoto, R.I. (1998) Genes Dev 12, 3788-96. 2 . Mercier, P.A. et al. (1999) J Cell Sci 112 ( Pt 16), 2765-74. 3 . Kim, S.A. et al. (2005) J Biol Chem 280, 12653-7. 4 . Holmberg, C.I. et al. (2001) EMBO J 20, 3800-10. 5 . Chu, B. et al. (1996) J Biol Chem 271, 30847-57. 6 . Wang, X. et al. (2003) Mol Cell Biol 23, 6013-26. 7 . Satyal, S.H. et al. (1998) Genes Dev 12, 1962-74. |
HeLa cells were either untreated (left panel) or heat shocked (right panel) for 1h. Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 cells and either 10 µl of HSF1 Antibody 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 HSPA6 Promoter Primers #5551, human HSP70 intron 1 primers, 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. HeLa细胞分为未处理（左侧）或热休克处理1小时（右侧）。使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003试剂盒，将4 x 10^6 个细胞来源的交联染色质，与10 μl HSF1 Antibody或者2 μl of Normal Rabbit IgG #2729进行染色质免疫沉淀。使用SimpleChIP® Human HSPA6 Promoter Primers #5551, 人HSP70内含子1引物和SimpleChIP® Human α Satellite Repeat Primers #4486进行real-time PCR来定量富集的DNA。input的染色质的量相当于1，每个样本中免疫沉淀的DNA的量在图中以相对于input的量来表示。 | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using HSF1 Antibody in the presence of control peptide (left) or antigen-specific peptide (right). 对石蜡包埋的人乳腺癌使用HSF1 Antibody进行免疫组织化学分析，其中左图为存在对照肽，右图为存在抗原特异性肽。 | |
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using HSF1 Antibody. 对石蜡包埋的人结肠癌使用HSF1 Antibody进行免疫组化分析。 | |
Immunohistochemical analysis of paraffin-embedded human pituitary adenoma, using HSF1 Antibody. 对石蜡包埋的人垂体腺瘤使用HSF1 Antibody进行免疫组化分析。 | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using HSF1 Antibody. 对石蜡包埋的人肺癌使用HSF1 Antibody进行免疫组化分析。 | |
Flow cytometric analysis of K562 cells, using HSF1 Antibody (blue) compared to a nonspecific negative control antibody (red). 使用HSF1抗体（蓝）和非特异的阴性对照抗体（红），对K562细胞进行流式细胞术分析。 | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma showing nuclear localization, using HSF1 Antibody. 对石蜡包埋的人乳腺癌使用HSF1抗体进行免疫组化分析，显示细胞核定位。 | |
DAPI staining (left) and immunofluorescent staining (right) of paraformaldehyde-fixed HeLa cells, using HSF1 antibody. 对多聚甲醛固定的HeLa细胞使用HSF1 antibody进行DAPI染色（左）和共聚焦免疫荧光染色（右）。 | |
Western blot analysis of extracts from HeLa, NIH/3T3, C6 and COS cells, using HSF1 antibody. 对HeLa, NIH/3T3, C6和COS细胞裂解液使用HSF1 antibody进行Western blot分析。 |
