Application
Immunofluorescent analysis of HSP27 in C6 cells. HSP27 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with HSP27 antibody [G3.1] at a dilution of 1:20 over night at 4?C, washed with PBS and incubated with a proper secondary antibody. Images were taken at 60X magnification.Application
Immunoprecipitation of HSP27 from HeLa cell lysate. Antigen-antibody complexes were formed by incubating 500μg whole cell lysate with 2μg of HSP27 antibody [G3.1] overnight on a rocking platform at 4°C. The immune complexes were captured on 50μl Protein A/G Agarose , washed extensively, and eluted. HeLa cell lysate (25μg) was loaded as a positive control. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with HSP27 antibody [G3.1] at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with a proper secondary antibody. Chemiluminescent detection was performed.Application
Western blot analysis of HSP27 in 50μg of various cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with HSP27 antibody [G3.1] at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a HRP-conjugated secondary antibody. Chemiluminescent detection was performed.| Product Name | HSP27 antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Mouse monoclonal antibody [G3.1] to HSP27The protein encoded by this gene is induced by environmental stress and developmental changes. The encoded protein is involved in stress resistance and actin organization and translocates from the cytoplasm to the nucleus upon stress induction. Defects in this gene are a cause of Charcot-Marie-Tooth disease type 2F (CMT2F) and distal hereditary motor neuropathy (dHMN). [provided by RefSeq, Oct 2008] |
| Immunogen | Partially purified human HSP27. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG1 |
| Clone Number | G3.1 |
| Host Species | Mouse |
| Tested Applications | ELISAICC/IFIHCIHC-PIMIPWB |
| IHC-P: 1:500: |
|
| Species Reactivity | HumanMouseDogRatPrimate |
| Concentration | 1mg/ml |
| Gene Synonyms | Hsp25 HSP27 heat shock 27kDa protein 1 HSPB1 HSP28 HS.76067 CMT2F HMN2B SRP27 |
|---|---|
| Target | HSP27/Heat Shock Protein 27 |
Application
Immunofluorescent analysis of HSP27 in C6 cells. HSP27 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with HSP27 antibody [G3.1] at a dilution of 1:20 over night at 4?C, washed with PBS and incubated with a proper secondary antibody. Images were taken at 60X magnification.
Application
Immunoprecipitation of HSP27 from HeLa cell lysate. Antigen-antibody complexes were formed by incubating 500μg whole cell lysate with 2μg of HSP27 antibody [G3.1] overnight on a rocking platform at 4°C. The immune complexes were captured on 50μl Protein A/G Agarose , washed extensively, and eluted. HeLa cell lysate (25μg) was loaded as a positive control. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with HSP27 antibody [G3.1] at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with a proper secondary antibody. Chemiluminescent detection was performed.
Application
Western blot analysis of HSP27 in 50μg of various cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with HSP27 antibody [G3.1] at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a HRP-conjugated secondary antibody. Chemiluminescent detection was performed.| Application Notes | IHC-P: 1:500: |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | -20° C, Avoid Freeze/Thaw Cycles |
| Storage Buffer | Ascites containing 0.05 % sodium azide |
