HSP27 Antibody [Unconjugated]

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.

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 the12-230 kDa separation system.

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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, KO
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

HSP27 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human HSP27
  Thr2-Lys205
  Accession # P04792
• Specificity: Detects human, mouse and rat HSP27.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: HSPB1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Knockout Validated 0.5 ug/mL
  • Simple Western 5 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for HSP27 Antibody [Unconjugated]

• 28 kDa heat shock protein
• DKFZp586P1322
• Estrogen-regulated 24 kDa protein
• Heat shock 27 kDa protein
• heat shock 27kD protein 1
• heat shock 27kDa protein 1
• heat shock protein beta-1
• HMN2B
• HS.76067
• HSP25
• HSP27
• HSP27HSP 27
• HSP28CMT2F
• HSPB1
• SRP27
• Stress-responsive protein 27

Background

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.

1. Gusev, N.B. et al. (2002) Biochemistry (Moscow) 67:511.
2. Garrido, C. et al. (2001) Biochem. Biophys. Res. Commun. 286:433.
3. Garrido, C. (2002) Cell Death Diffr. 9:483.
4. Brvey, J-M. et al. (2000) Nat. Cell Biol. 2:645.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HSP27 Antibody (AF15801)(3)

Publications using AF15801

• Wellslager, B;Roberts, J;Chowdhury, N;Madan, L;Orellana, E;Yilmaz, Ö; Porphyromonas gingivalis activates Heat-Shock-Protein 27 to drive a LC3C-specific probacterial form of select autophagy that is redox sensitive for intracellular bacterial survival in human gingival mucosa bioRxiv : the preprint server for biology 2024-07-02 [PMID: 39005460] (Immunocytochemistry, Human)
• Vanessa Fétaud-Lapierre, Catherine M. Pastor, Annarita Farina, Denis F. Hochstrasser, Jean-Louis Frossard, Pierre Lescuyer Proteomic Analysis of Heat Shock-Induced Protection in Acute Pancreatitis Journal of Proteome Research 11/5/2010 [PMID: 20815342]
• Lin YP, Hsu ME, Chiou YY Comparative proteomic analysis of rat aorta in a subtotal nephrectomy model. Proteomics, 2010-07-01;10(13):2429-43. 2010-07-01 [PMID: 20405472] (IHC-P, Western Blot, Rat)

Bioinformatics

• Gene Symbol: HSPB1
• Uniprot:
  • P04792()