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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human, Mouse, Rat, Bovine, Canine, Guinea Pig, Hamster, Sheep

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockout Validated, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 8A7

Concentration

1 mg/ml

Product Specifications

Immunogen

HSP27 peptide

Localization

Cytoplasm , Nucleus , Cytoskeleton , Spindle

Specificity

Detects approx 25kDa or approx 27kDa. Recognizes HSP25 and HSP27, cross reacts with alpha B crystallin.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for HSP27 Antibody (8A7)

Knockout Validated: HSP27 Antibody (8A7) [NB110-96431]

Knockout Validated: HSP27 Antibody (8A7) [NB110-96431]

Knockout Validated: HSP27 Antibody (8A7) [NB110-96431] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HSP27 knockout (KO) HeLa cell line. PVDF membrane was probed with 1:8,000 of Mouse Anti-Human HSP27 Monoclonal Antibody (Catalog # NB110-96431) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (catalog number HAF018). Specific band was detected for HSP27 at approximately 27 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.
Western Blot: HSP27 Antibody (8A7) [NB110-96431]

Western Blot: HSP27 Antibody (8A7) [NB110-96431]

Western Blot: HSP27 Antibody (8A7) [NB110-96431] - analysis of Rat Lung tissue lysates showing detection of Hsp27 protein using Mouse Anti-Hsp27 Monoclonal Antibody, Clone 8A7 . Load: 15 ug protein. Block: 5% blocking solution. Primary Antibody: Mouse Anti-Hsp27 Monoclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse: HRP for 1 hour at RT.
Immunocytochemistry/ Immunofluorescence: HSP27 Antibody (8A7) [NB110-96431]

Immunocytochemistry/ Immunofluorescence: HSP27 Antibody (8A7) [NB110-96431]

Immunocytochemistry/Immunofluorescence: HSP27 Antibody (8A7) [NB110-96431] - analysis using Mouse Anti-Hsp27 Monoclonal Antibody, Clone 8A7 . Tissue: embryo somites. Species: Rat. Primary Antibody: Mouse Anti-Hsp27 Monoclonal Antibody at 1:1000. Secondary Antibody: FITC Goat Anti-Mouse (green). Counterstain: Rhodamine-phalloidin labeled actin (red). Courtesy of: Mike Welsh, Umich.

Applications for HSP27 Antibody (8A7)

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:200

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Western Blot

1:5000
Application Notes
A 1:5000 dilution of HSP25/HSP27 Antibody was sufficient for detection of HSP27 in 20 ug of HeLa cell lysate by ECL immunoblot analysis.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS, 50% Glycerol

Preservative

0.09% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: HSP27

Hsp25 is the mouse homologue of the human Hsp27 protein, a member of the small heat shock protein family comprised of a diverse group of proteins from approx. 15 to >30kDa(1). The basic structure of most sHsps is a homologous and highly conserved amino acid sequence, with an alpha-crystallin-domain at the C-terminus and the WD/EPF domain at the less conserved N-terminus. This N-terminus is essential for the development of high molecular oligomers (2, 3). Hsp27-oligomers consist of stable dimers formed by as many as 8-40 Hsp27 protein monomers (4). The oligomerization status is connected with the chaperone activity: aggregates of large oligomers have high chaperone activity, whereas dimers have no chaperone activity (5). HSP27 is localized to the cytoplasm of unstressed cells but can redistribute to the nucleus in response to stress, where it may function to stabilize DNA and/or the nuclear membrane. It can be rapidly phosphorylated in response to physiological stimuli relevant to the cell type examined. Thus, hsp27 has been suggested to be an important intermediate in second messenger mediated signaling pathways (6). Other functions include chaperone activity (as mentioned above), thermotolerance in vivo, inhibition of apoptosis, and signal transduction. Specifically, in vitro, it acts as an ATP-independent chaperone by inhibiting protein aggregation and by stabilizing partially denatured proteins, which ensures refolding of the HSP70 complex. Hsp27 is also involved in the apoptotic signaling pathway because it interferes with the activation of cytochrome c/Apaf-1/dATP complex, thereby inhibiting the activation of procaspase-9. It is also hypothesized that hsp27 may serve some role in cross-bridge formation between actin and myosin (7). And finally, Hsp27 is also thought to be involved in the process of cell differentiation. The up-regulation of Hsp27 correlates with the rate of phosphorylation and with an increase of large oligomers. It is possible that Hsp27 may play a crucial role in termination of growth (8).

Long Name

Heat Shock Protein 27

Alternate Names

HSP25, HSPB1

Entrez Gene IDs

3315 (Human); 15507 (Mouse); 24471 (Rat)

Gene Symbol

HSPB1

UniProt

Additional HSP27 Products

Product Documents for HSP27 Antibody (8A7)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for HSP27 Antibody (8A7)

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.

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