Phospho-Hsp27 (S78) Recombinant Rabbit Monoclonal Antibody [JJ08-70]
cat.: ET1701-19
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, IP |
| Clonality: | Monoclonal |
| Clone number: | JJ08-70 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 23 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser78 of human Hsp27. |
| Positive control: | Human heart tissue lysate, human skeletal muscle tissue lysate, Hela, MCF-7, NIH/3T3, human breast carcinoma tissue, Hela cell lysate, Hela cell lysate treated with UV for 2 hours, human cervical carcinoma tissue. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P IP |
1:1,000-1:2,000 1:50-1:200 1:50-1:200 1:50-1:400 Use at an assay dependent concentration. |
| Uniprot #: | SwissProt: P04792 Human |
| Alternative names: | Heat shock 27kDa protein 28 kDa heat shock protein CMT2F DKFZp586P1322 epididymis secretory protein Li 102 Estrogen regulated 24 kDa protein Estrogen-regulated 24 kDa protein Heat shock 25kDa protein 1 Heat shock 27 kDa protein Heat shock 27kD protein 1 Heat shock 27kDa protein 1 Heat shock 28kDa protein 1 Heat Shock Protein 27 Heat shock protein beta 1 Heat shock protein beta-1 heat shock protein family B (small) member 1 HEL-S-102 HMN2B HS.76067 Hsp 25 HSP 27 Hsp 28 Hsp B1 Hsp25 HSP27 Hsp28 HspB1 HSPB1_HUMAN SRP27 Stress responsive protein 27 Stress-responsive protein 27 |
Images
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Fig1:
Western blot analysis of Phospho-Hsp27 (S78) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-19, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Human heart tissue lysate Lane 2: Human skeletal muscle tissue lysate Predicted band size: 23 kDa Observed band size: 27 kDa |
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Fig2: ICC staining of Phospho-Hsp27 (S78) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-19, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). |
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Fig3: ICC staining of Phospho-Hsp27 (S78) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-19, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). |
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Fig4: ICC staining of Phospho-Hsp27 (S78) in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-19, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). |
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Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-Hsp27 (S78) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-19, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig6:
Western blot analysis of Phospho-Hsp27 (S78) on different lysates with Rabbit anti-Phospho-Hsp27 (S78) antibody (ET1701-19) at 1/500 dilution. Lane 1: Hela cell lysate Lane 2: Hela cell lysate treated with UV for 2 hours Lysates/proteins at 10 µg/Lane. Predicted band size: 23 kDa Observed band size: 27 kDa Exposure time: 2 minutes; 15% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-19) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. |
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Fig7:
Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue with Rabbit anti-Phospho-Hsp27 (S78) antibody (ET1701-19) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-19) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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