Cell Signaling Technology

Product Pathways - Protein Folding

HSF1 Antibody #4356

heat shock   heat shock protein   Heat Shock Transcription Factor 1 (HSTF1)   hsf   hsf1  

No. Size Price
4356S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
4356T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价
4356 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 82 Rabbit
IP 1:50
IHC-P 1:250
F 1:50
IF-IC 1:500
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

This antibody detects endogenous levels of total HSF1 protein.

The antibody does not cross-react with other HSF proteins.

此抗体识别内源性的HSF1总蛋白。此抗体不与其它HSF蛋白相互作用。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids at the carboxy-terminus of human HSF1 protein. Antibodies are purified by protein A and peptide affinity chromatography.

该多克隆抗体用与人类HSF1蛋白羧基末端附近的氨基酸链对应的人工合成肽段免疫动物而制成。该抗体使用蛋白A和蛋白亲和层析纯化而得。

Western Blotting

Western Blotting

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。

IF-IC

IF-IC

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)。

Flow Cytometry

Flow Cytometry

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。

IHC-P (paraffin)

IHC-P (paraffin)

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。

IHC-P (paraffin)

IHC-P (paraffin)

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。

IHC-P (paraffin)

IHC-P (paraffin)

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。

IHC-P (paraffin)

IHC-P (paraffin)

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。

IHC-P (paraffin)

IHC-P (paraffin)

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。

Chromatin IP

Chromatin IP

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。

Background

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).

所有生物都通过快速诱导高表达保守的热休克蛋白(HSP)来应对温度增加和其它环境应激,它能够将失活的蛋白去折叠,也能促进损坏蛋白的降解。热休克基因转录被热休克因子(HSF)家族和转录激活因子调节,它们可以结合到所有热休克基因上游的热反应元件(HSE)(1)。HSE在各生物中是高度保守的,包括多个相邻和反向的五核苷酸重复序列基序5'-nGAAn-3'。HSF的保守性稍差,大概共享40%的序列。脊椎动物细胞包括四种HSF蛋白:HSF1、2和4是普遍存在的,HSF3只在鸟类中存在。HSF1在应对热、重金属和氧化物质时诱导热休克基因转录,HSF2与生精功能和胚胎发育有关。HSF3和HSF4的功能与HSF1和HSF2有重叠。失活的HSF1作为单体存在,定位于细胞质和细胞核,但不与DNA结合(1,2)。应激时,HSF1磷酸化,形成同源三聚体,结合到DNA上级或热休克基因转录(1,2)。HSF1活性由PLK对其Ser419的磷酸化所正向调节,可以增加其核转位,也能由CaMKII对其Ser230磷酸化,加强反式激活(3,4)。另外,HSF1活性能够被GSK3和ERK1分别对其Ser303和Ser307磷酸化而抑制,从而导致HSF1与14-3-3蛋白结合并被隔离在细胞质中(5,6)。在热休克反应衰减时,HSF1被HSP70、HSP40/Hdj-1和HSF结合蛋白1(HSBP1)直接结合抑制(7)。

  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.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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