GTX25478

antibody from GeneTex

Targeting: HSPD1 GroEL, HSP60, SPG13

Western blot (Supportive data in Antibodypedia)

ELISA (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Data presented on provider website)

Blocking/Neutralizing (Data presented on provider website)

Antibody Data

Product number

GTX25478

Provider

GeneTex

Proper citation

GeneTex Cat#GTX25478, RRID:AB_373944

Product name

HSP60 antibody [4B9/89]

Antibody type

Monoclonal

Reactivity

Human, Mouse, Rat, Bacteria, Simian

Host

Mouse

Western blot

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Western blot analysis of HSP60 in 50£gg 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¢XC on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a proper secondary antibody. Chemiluminescent detection was performed.

Immunocytochemistry

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

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.

Immunoprecipitation

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Immunoprecipitation of HSP60 from HeLa cells. Antigen-antibody complexes were formed by incubating 500£gg whole cell lysate with 2£gg of HSP60 antibody [4B9/89] overnight on a rocking platform at 4¢XC. The immune complexes were captured on 50£gl 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¢XC, washed in TBST, and probed with a proper secondary antibody. Chemiluminescent detection was performed.

Immunohistochemistry

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

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¢XC 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.