Human HSP27 Antibody

R&D Systems | Catalog # AF1580

R&D Systems
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Knockout Validated, Western Blot, Simple Western

Cited:

Western Blot, Flow Cytometry, Simple Western

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all HSP27 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human HSP27
Accession # P04792

Specificity

Detects recombinant and endogenous human HSP27 in Western blots.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human HSP27 Antibody

Detection of Human HSP27 by Western Blot.

Western Blot shows lysates of HeLa human cervical epithelial carcinoma cell line and DU145 human prostate carcinoma cell line. PVDF membrane was probed with 0.1 µg/ml of Rabbit Anti-Human HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1580) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for HSP27 at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
Detection of Human HSP27 antibody by Simple WesternTM.

Detection of Human HSP27 by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for HSP27 at approximately 31 kDa (as indicated) using 0.5 µg/mL of Rabbit Anti-Human HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1580). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western Blot Shows Human HSP27 Antibody Specificity by Using Knockout Cell Line.

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 Rabbit Anti-Human HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1580) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). 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 (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of HSP27 by Western Blot

Detection of HSP27 by Western Blot

Phosphorylation of heat shock protein 27 (HSP27) after mechanical strain with different force magnitudes for different time spans (a). Densitometric analysis of HSP27 phosphorylation in relation to HSP27 expression after exposing human periodontal ligament fibroblasts (PDLF) to different force magnitudes for 6 h (b). Statistics: Welch analysis of variance (ANOVA) with Dunnett’s T3 multiple comparisons tests; *P < 0.05Phosphorylierung von HSP27 (Hitzeschockprotein 27) nach mechanischer Belastung mit unterschiedlichen Kraftstärken für verschiedene Zeitspannen (a). Densitometrische Analyse der HSP27-Phosphorylierung im Verhältnis zur HSP27-Expression nach 6‑stündiger Exposition von PDLF (parodontale Ligamentfibroblasten) mit unterschiedlichen Kraftstärken (b). Statistik: Welch-ANOVA („analysis of variance“) mit Dunnett-T3 Post-hoc-Test; *p < 0,05 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35445818), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HSP27 by Western Blot

Detection of HSP27 by Western Blot

Heat shock protein 27 (HSP27) phosphorylation was upregulated after compression and transfection of pHSP27+ (a). Transfection of pHSP27+ reduced the cell number (b) and increased lactate dehydrogenase (LDH) release (c) in periodontal ligament fibroblasts (PDLF) without and with compression. Statistics: Welch analysis of variance (ANOVA) with Dunnett’s T3 multiple comparisons tests; *P < 0.05; **P < 0.01; ***P < 0.001Die HSP27(Hitzeschockprotein 27)-Phosphorylierung wurde nach Kompression und Transfektion von pHSP27+ hochreguliert (a). Die Transfektion von pHSP27+ reduzierte die Zellzahl (b) und erhöhte die LDH(Laktatdehydrogenase)-Freisetzung (c) in PDLF (parodontale Ligamentfibroblasten) ohne und mit Kompression. Statistik: Welch-ANOVA („analysis of variance“) mit Dunnett-T3 Post-hoc-Test; *p < 0,05; **p < 0,01; ***p < 0,001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35445818), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human 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

0.5 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

0.1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and DU145 human prostate carcinoma cell line

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.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*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

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: HSP27

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. Full length human HSP27 shares 83% and 81% aa identity with mouse and rat HSP27, respectively.

References

  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.

Long Name

Heat Shock Protein 27

Alternate Names

HSP25, HSPB1

Entrez Gene IDs

3315 (Human); 15510 (Mouse); 24471 (Rat)

Gene Symbol

HSPB1

UniProt

P04792

Additional HSP27 Products

Product Documents for Human HSP27 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human HSP27 Antibody

For research use only

Citations for Human HSP27 Antibody

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