WB result of Phospho-HSP27 (Ser82) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody incubation conditions: overnight at 4°C
Primary antibody: Phospho-HSP27 (Ser82) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 25 μg/ml Anisomycin for 30 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 23 kDa
Observed MW: 25 kDa
Phospho-HSP27 (Ser82) Recombinant Rabbit mAb (S-4792)
Phospho-HSP27 (Ser82) Recombinant Rabbit mAb (S-4792)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-HSP27 (Ser82) |
| Synonyms | Heat shock protein beta-1; 28 kDa heat shock protein; Estrogen-regulated 24 kDa protein; Heat shock 27 kDa protein (HSP 27); Heat shock protein family B member 1; Stress-responsive protein 27 (SRP27); HSP27; HSP28; HSPB1 |
| Location | Cytoplasm, Cytoskeleton, Nucleus |
| Accession | P04792 |
| Clone Number | S-4792 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms |
| Positive Sample | HeLa treated with 25 μg/ml Anisomycin for 30 minutes, C2C12 treated with 25 μg/ml Anisomycin for 30 minutes |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms |
| IHC-P | 1:100 | Hu |
| ICC | 1:50-1:100 | Hu, Ms |
Background
Phospho-HSP27 (Ser82) refers to the heat shock protein 27 (HSP27, also known as HSPB1) specifically phosphorylated at the serine 82 residue, a critical post-translational modification primarily mediated by the MAPKAP kinase 2 (MK2) downstream of the p38 MAPK signaling pathway in response to cellular stressors such as heat shock, oxidative stress, or inflammatory cytokines. This specific phosphorylation event induces a conformational change that causes the dissociation of large HSP27 oligomers into smaller dimers or monomers, thereby enhancing its chaperone activity and ability to interact with client proteins, while simultaneously playing a pivotal role in stabilizing the actin cytoskeleton, inhibiting apoptosis by preventing cytochrome c release and caspase activation, and promoting cell survival and proliferation. Due to its significant upregulation and activation in various malignancies, Phospho-HSP27 (Ser82) is widely recognized not only as a key regulator of cellular stress responses and cytoprotection but also as a potential biomarker for cancer prognosis and a therapeutic target in diseases characterized by dysregulated stress signaling pathways.
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Western Blot
WB result of Phospho-HSP27 (Ser82) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody incubation conditions: overnight at 4°C
Primary antibody: Phospho-HSP27 (Ser82) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated C2C12 whole cell lysate 20 µg
Lane 2: C2C12 treated with 25 μg/ml Anisomycin for 30 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 23 kDa
Observed MW: 25 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cervical squamous cell carcinoma. Anti- Phospho-HSP27 (Ser82) antibody was used at 1/100 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows negative staining in lambda phosphatase treated paraffin-embedded human cervical squamous cell carcinoma. Anti- Phospho-HSP27 (Ser82) antibody was used at 1/100 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti- Phospho-HSP27 (Ser82) antibody was used at 1/100 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows negative staining in lambda phosphatase treated paraffin-embedded human colon cancer. Anti- Phospho-HSP27 (Ser82) antibody was used at 1/100 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human tonsil. Anti- Phospho-HSP27 (Ser82) antibody was used at 1/100 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows negative staining in lambda phosphatase treated paraffin-embedded human tonsil. Anti- Phospho-HSP27 (Ser82) antibody was used at 1/100 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC analysis of Hela cells treated with λ phosphatase (top panel) and untreated Hela cells (below panel). Anti- Phospho-HSP27 (Ser82) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ICC analysis of C2C12 cells treated with λ phosphatase (top panel) and untreated C2C12 cells (below panel). Anti- Phospho-HSP27 (Ser82) antibody was used at 1/50 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
