Heat shock proteins (HSPs) are a family of highly conserved stress response proteins. Heat shock proteins function primarily as molecular chaperones by facilitating the folding of other cellular proteins, preventing protein aggregation or targeting improperly folded proteins to specific degradative pathways. HSPs are typically expressed at low levels under normal physiological conditions but are dramatically up-regulated in response to cellular stress. Elevated levels of HSPs have been observed in association with ischemia/reperfusion, cancer, and chronic heart failure. HSP27, also known as HSPB1, is a member of the small heat shock protein family, which also includes HSP25 and the alpha -crystallins. HSP27 forms a large oligomer and the extent of phosphorylation plays a role in determining specific functions. HSP27 also functions as an anti-apoptotic molecule, regulating apoptosis through direct interaction with key components of the apoptotic pathway. HSP27 binds and sequesters cytochrome c released from the mitochondria in response to an apoptotic stimulus. This prevents the proper assembly of the apoptosome and subsequently, the activation of procaspase-9 and procaspase-3.
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Key Product Details
Validated by
Knockout/Knockdown, Biological Validation
Species Reactivity
Validated:
Human, Mouse, Rat
Cited:
Human, Rat
Applications
Validated:
Knockout Validated, Western Blot, Simple Western
Cited:
Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human HSP27
Thr2-Lys205
Accession # P04792
Thr2-Lys205
Accession # P04792
Specificity
Detects human, mouse and rat HSP27.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human/Mouse/Rat HSP27 Antibody
Detection of Human and Mouse HSP27 by Western Blot.
Western blot shows lysates of Jurkat human acute T cell leukemia cell line and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with a 42 °C heat shock for 30 minutes with a 3 hour recovery. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human/Mouse/Rat HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF15801), followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for HSP27 at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Human HSP27 by Simple WesternTM.
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for HSP27 at approximately 33 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF15801) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western Blot Shows Human HSP27 Specificity by Using Knockout Cell Line.
Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HSP27 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF15801) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for HSP27 at approximately 27 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control.This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Applications for Human/Mouse/Rat HSP27 Antibody
Application
Recommended Usage
Knockout Validated
HSP27 is specifically detected in HeLa human
cervical epithelial carcinoma parental cell line but is not detectable in
HSP27 knockout HeLa cell line.
Simple Western
5 µg/mL
Sample: Jurkat human acute T cell leukemia cell line
Sample: Jurkat human acute T cell leukemia cell line
Western Blot
0.1 µg/mL
Sample: 42° C heat shock-treated Jurkat human acute T cell leukemia cell line and NIH‑3T3 mouse embryonic fibroblast cell line
Sample: 42° C heat shock-treated Jurkat human acute T cell leukemia cell line and NIH‑3T3 mouse embryonic fibroblast cell line
Reviewed Applications
Read 2 reviews rated 4 using AF15801 in the following applications:
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: HSP27
References
- Gusev, N.B. et al. (2002) Biochemistry (Moscow) 67:511.
- Garrido, C. et al. (2001) Biochem. Biophys. Res. Commun. 286:433.
- Garrido, C. (2002) Cell Death Diffr. 9:483.
- Brvey, J-M. et al. (2000) Nat. Cell Biol. 2:645.
Long Name
Heat Shock Protein 27
Alternate Names
HSP25, HSPB1
Gene Symbol
HSPB1
UniProt
Additional HSP27 Products
Product Documents for Human/Mouse/Rat HSP27 Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human/Mouse/Rat HSP27 Antibody
For research use only
Related Research Areas
Citations for Human/Mouse/Rat HSP27 Antibody
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4 out of 5
2 Customer Ratings
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Application: MicroarraysSample Tested: EDTA PlasmaSpecies: HumanVerified Customer | Posted 11/20/2018
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
