Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 1026B
View all HSP27 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing the human HSP27 S78/S82 site
Accession # P04792

Specificity

Detects human HSP27 when dually phosphorylated at S78/S82 in Western blots.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human phospho-HSP27 (S78/S82) Antibody (1026B)

Detection of Human Phospho-HSP27 (S78/S82) antibody by Western Blot.

Detection of Human Phospho-HSP27 (S78/S82) by Western Blot.

Western blot shows lysates of MCF-7 human breast cancer cell line and HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 20 mJ/cm2ultraviolet light (UV) with a 30 minute recovery. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human Phospho-HSP27 (S78/S82) Monoclonal Antibody (Catalog # MAB23141) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-HSP27 (S78/S82) at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-HSP27 (S78/S82) antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

Phospho-HSP27 (S78/S82) in HeLa Human Cell Line.

HSP27 phosphorylated at S78/S82 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line unstimulated (lower panel) or stimulated with 20 mJ/cm2ultraviolet radiation (upper panel) using Rabbit Anti-Human Phospho-HSP27 (S78/S82) Monoclonal Antibody (Catalog # MAB23141) at a 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Phospho-HSP27 (S78/S82) by Western Blot

Detection of Phospho-HSP27 (S78/S82) by Western Blot

Phosphorylation of Akt, RSK, and CREB during T. cruzi infection. Lysates from primary HCoEpiC infected with T. cruzi at multiple time points were resolved by SDS-PAGE, blotted, and probed with antibodies against (A) p-HSP27 (B) p-Akt T308, (C) p-Akt S473, (D) p-RSK, and (E) p-CREB, and developed as described. The blots were stripped and reprobed with antibodies against the respective total proteins. The blots were developed by chemiluminescence and scanned. The normalized fold change in the level of each phosphorylated protein was determined and plotted in the bar graph below the respective blot. The bar graphs represent mean values ± SE from three independent biological replicates. The value of p<0.05 was considered significant; *p<0.05 **p<0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30222739), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human phospho-HSP27 (S78/S82) Antibody (1026B)

Application
Recommended Usage

Immunocytochemistry

5-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line unstimulated or stimulated with 20 mJ/cm2 ultraviolet radiation

Western Blot

1 µg/mL
Sample: MCF‑7 human breast cancer cell line and HeLa human cervical epithelial carcinoma cell line treated with ultraviolet light (UV)

Reviewed Applications

Read 1 review rated 5 using MAB23141 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 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. *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

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

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 is a member of the small heat shock protein family, which also includes HSP25 and the alpha -crystallins. Ser78 and Ser82 of HSP27 are phosphorylated in vivo in response to growth factors or heat shock, and the extent of phosphorylation plays a role in determining specific functions. HSP27 forms a large oligomer and 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.

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 phospho-HSP27 (S78/S82) Antibody (1026B)

Certificate of Analysis

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Product Specific Notices for Human phospho-HSP27 (S78/S82) Antibody (1026B)

For research use only

Citations for Human phospho-HSP27 (S78/S82) Antibody (1026B)

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  • Human Phospho-HSP27 (S78/S82) Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HeLa cells
    Species: Human
    Verified Customer | Posted 05/17/2022
    Human phospho-HSP27 (S78/S82) Antibody (1026B) MAB23141

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