Product Pathways - Protein Folding
HSF1 (D3L8I) Rabbit mAb #12972
|12972S||100 µl (10 western blots)||---||In Stock||---|
|12972||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat, Monkey, Bovine, Dog, Pig||Endogenous||80||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP
Specificity / Sensitivity
HSF1 (D3L8I) Rabbit mAb recognizes endogenous levels of total HSF1 protein. This antibody is not predicted to cross-react with HSF2.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human HSF1 protein.
Western blot analysis of extracts from various cell lines using HSF1 (D3L8I) Rabbit mAb.
Immunoprecipitation of HSF1 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2), HSF1 Antibody #4356 (lane 3) or HSF1 (D8L8I) Rabbit mAb (lane 4). Lane 1 is 10% input. Western blot analysis was performed using HSF1 (D8L8I) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using HSF1 (D3L8I) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using HSF1 (D3L8I) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using HSF1 (D3L8I) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded mouse spleen, HSF1 wild type (left), HSF1 knock out (middle) or HSF2 knock out (right), using HSF1 (D3L8I) Rabbit mAb. Tissues courtesy of Dr. Nahid Mivechi, Georgia Health Sciences University, Atlanta, Georgia.
Flow cytometric analysis of HeLa cells using HSF1 (D3L8I) Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). 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, untreated (upper) or heat-treated (43.5ºC, 1 hr without recovery; lower), using HSF1 (D3L8I) Rabbit mAb (green). Actin filaments were labeled with DyLight® 554 Phalloidin #13054 (red). HSF1 is redistributed to nuclear stress granules following heat shock (insets).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells heat shocked for 1h and either 10 µl of HSF1 (D3L8I) 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 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.
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 that 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 Ser419 by PLK1, which enhances nuclear translocation, and phosphorylation of Ser230 by CaMKII, which enhances transactivation (3,4). Alternatively, HSF1 activity is repressed by phosphorylation of serines at 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).
- Morimoto, R.I. (1998) Genes Dev 12, 3788-96.
- Mercier, P.A. et al. (1999) J Cell Sci 112 ( Pt 16), 2765-74.
- Kim, S.A. et al. (2005) J Biol Chem 280, 12653-7.
- Holmberg, C.I. et al. (2001) EMBO J 20, 3800-10.
- Chu, B. et al. (1996) J Biol Chem 271, 30847-57.
- Wang, X. et al. (2003) Mol Cell Biol 23, 6013-26.
- Satyal, S.H. et al. (1998) Genes Dev 12, 1962-74.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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