Application
Immunoprecipitation of HSP60 from HeLa cells. Antigen-antibody complexes were formed by incubating 500μg whole cell lysate with 2μg of HSP60 antibody [4B9/89] overnight on a rocking platform at 4°C. The immune complexes were captured on 50μl Protein A/G Agarose , washed extensively, and eluted. Samples were then 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 HSP60 antibody [4B9/89] 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
Immunohistochemistry was performed on normal deparaffinized human Kidney 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 1:100 with or without HSP60 antibody [4B9/89] 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
Immunofluorescent analysis of HSP60 in A2058 cells. HSP60 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 HSP60 antibody [4B9/89] at a dilution of 1:200 over night at 4?C, washed with PBS and incubated with a proper secondary antibody. Images were taken at 60X magnification.Application
Western blot analysis of HSP60 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 HSP60 antibody [4B9/89] 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 | HSP60 antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Mouse monoclonal antibody [4B9/89] to HSP60This gene encodes a member of the chaperonin family. The encoded mitochondrial protein may function as a signaling molecule in the innate immune system. This protein is essential for the folding and assembly of newly imported proteins in the mitochondria. This gene is adjacent to a related family member and the region between the 2 genes functions as a bidirectional promoter. Several pseudogenes have been associated with this gene. Two transcript variants encoding the same protein have been identified for this gene. Mutations associated with this gene cause autosomal recessive spastic paraplegia 13. [provided by RefSeq, Jun 2010] |
| Immunogen | Human placental HSP60. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG2a |
| Clone Number | 4B9/89 |
| Host Species | Mouse |
| Tested Applications | BlockingELISAICC/IFIHCIHC-PIPWB |
| ICC/IF: 1:50: |
|
| Species Reactivity | HumanMouseRatHamsterPrimateBacteria |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Synonyms | HSPD1 HuCHA60 SPG13 CPN60 HSP65 HSP-60 GROEL HSP60 heat shock 60kDa protein 1 (chaperonin) HLD4 |
|---|---|
| Target | HSP60/Heat Shock Protein 60 |
Application
Immunoprecipitation of HSP60 from HeLa cells. Antigen-antibody complexes were formed by incubating 500μg whole cell lysate with 2μg of HSP60 antibody [4B9/89] overnight on a rocking platform at 4°C. The immune complexes were captured on 50μl Protein A/G Agarose , washed extensively, and eluted. Samples were then 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 HSP60 antibody [4B9/89] 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
Immunohistochemistry was performed on normal deparaffinized human Kidney 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 1:100 with or without HSP60 antibody [4B9/89] 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
Immunofluorescent analysis of HSP60 in A2058 cells. HSP60 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 HSP60 antibody [4B9/89] at a dilution of 1:200 over night at 4?C, washed with PBS and incubated with a proper secondary antibody. Images were taken at 60X magnification.
Application
Western blot analysis of HSP60 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 HSP60 antibody [4B9/89] 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: 1:50: |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | -20° C, Avoid Freeze/Thaw Cycles |
| Storage Buffer | phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. |
