Immunofluorescent analysis of TRAP1 in NCI-H460 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a TRAP1 monoclonal antibody (Product # MA1-010) at a dilution of 1:200 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). TRAP1 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Mouse|
|Immunogen||Purified recombinant TRAP1.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:20|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:2,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-010 detects tumor necrosis factor receptor-associated protein (TRAP1) from human tissues.
MA1-010 has been successfully used in Western blot, immunofluorescence and immunoprecipitation procedures. By Western blot, this antibody detects an ~75 kDa protein representing TRAP1. Immunofluorescence staining of TRAP1 in PC-3-M cells with MA1-010 produces a pattern consistent with mitochondrial staining. Immunoprecipitation of TRAP1 using MA1-010 fails to co-precipitate p23, Hop, or CyP40 suggesting TRAP1 and quote;s inability to associate with these co-chaperones.
The MA1-010 immunogen is purified, recombinant, human TRAP1.
The 90 kDa heat shock protein (hsp90) family of molecular chaperones is a highly conserved family of proteins that play an important physiological role. Hsp90 is involved in numerous cellular processes but is best known for its association with signal transduction machinery. A recently cloned homolog of hsp90 is the tumor necrosis factor receptor-associated protein (TRAP1). Like hsp90, TRAP1 is found to be associated with numerous proteins involved in diverse actions. Immunofluorescence data has shown TRAP1 to be localized in the mitochondria of mammalian cells. This observation and the fact that TRAP1 is shown to have a mitochondrial targeting presequence strongly implicates TRAP1 as a mitochondrial matrix protein.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Cytotoxicity of withaferin A in glioblastomas involves induction of an oxidative stress-mediated heat shock response while altering Akt/mTOR and MAPK signaling pathways.
MA1-010 was used in western blot to study the molecular mechanism of withaferin A cytotoxicity in glioblastomas
|Grogan PT,Sleder KD,Samadi AK,Zhang H,Timmermann BN,Cohen MS||Investigational new drugs (31:545)||2013|
A novel C-terminal HSP90 inhibitor KU135 induces apoptosis and cell cycle arrest in melanoma cells.
MA1-010 was used in western blot to study the effect of a C-terminal HSP90 inhibitor KU135 on melanoma cell death and proliferation
|Samadi AK,Zhang X,Mukerji R,Donnelly AC,Blagg BS,Cohen MS||Cancer letters (312:158)||2011|
Involvement of tumor necrosis factor receptor-associated protein 1 (TRAP1) in apoptosis induced by beta-hydroxyisovalerylshikonin.
MA1-010 was used in western blot to study the role of mitochondrial TRAP1 expression in the ROS-induced apoptosis.
|Masuda Y,Shima G,Aiuchi T,Horie M,Hori K,Nakajo S,Kajimoto S,Shibayama-Imazu T,Nakaya K||The Journal of biological chemistry (279:42503)||2004|
Hsp90 chaperone activity requires the full-length protein and interaction among its multiple domains.
MA1-010 was used in immunoprecipitation to investigate the regulation of Hsp90 function in vitro and in vivo
|Johnson BD,Chadli A,Felts SJ,Bouhouche I,Catelli MG,Toft DO||The Journal of biological chemistry (275:32499)||2000|