Application
Immunofluorescent analysis of HSP70 in HeLa cells. Cells were grown on slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with HSP70 antibody [4G4] at a dilution of 1:100 overnight at 4°C , washed with PBS and incubated with a proper secondary antibody. HSP70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.Application
Immunohistochemistry was performed on normal biopsies of deparaffinized human breast tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with or without HSP70 antibody [4G4] overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.Application
Western blot analysis of HSP70 in 50ug 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 HSP70 antibody [4G4] 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 proper secondary antibody. Chemiluminescent detection was performed.| Product Name | HSP70 antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Mouse monoclonal antibody [4G4] to HSP70This intronless gene encodes a 70kDa heat shock protein which is a member of the heat shock protein 70 family. In conjuction with other heat shock proteins, this protein stabilizes existing proteins against aggregation and mediates the folding of newly translated proteins in the cytosol and in organelles. It is also involved in the ubiquitin-proteasome pathway through interaction with the AU-rich element RNA-binding protein 1. The gene is located in the major histocompatibility complex class III region, in a cluster with two closely related genes which encode similar proteins. [provided by RefSeq, Jul 2008] |
| Immunogen | Human recombinant HSP70 over expressed in E. coli. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG1 |
| Clone Number | 4G4 |
| Host Species | Mouse |
| Tested Applications | BlockingFACSICC/IFIHCIHC-PInhibitionIPWB |
| ICC/IF: 1:100: |
|
| Species Reactivity | HumanMousePrimate |
| Concentration | 1mg/ml |
| Gene Synonyms | HSPA1 HSP70-1A HSP70-1 heat shock 70kDa protein 1A HSP70I HSP72 HSPA1A |
|---|---|
| Target | HSP70/Heat Shock Protein 70 |
| Tissue Specificity | This antibody specifically detects HSP70 and does not cross-react with other members of the HSP70 family. |
Application
Immunofluorescent analysis of HSP70 in HeLa cells. Cells were grown on slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with HSP70 antibody [4G4] at a dilution of 1:100 overnight at 4°C , washed with PBS and incubated with a proper secondary antibody. HSP70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
Application
Immunohistochemistry was performed on normal biopsies of deparaffinized human breast tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with or without HSP70 antibody [4G4] overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Application
Western blot analysis of HSP70 in 50ug 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 HSP70 antibody [4G4] 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 proper secondary antibody. Chemiluminescent detection was performed.| Application Notes | ICC/IF: 1:100: |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | -20° C, Avoid Freeze/Thaw Cycles |
| Storage Buffer | Ascites |
