Anti-TRAP1 antibody [TRAP1-6]

• Supplier: Abcam
• Catalog: ab2721
• Prices: $390.00
• Sizes: 100 µg
• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG1
• Clone: TRAP1-6
• Applications: IHC-P IP WB IHC-F ICC/IF ICC/IF
• Species Reactivities: Mouse, Human
• Antigen: Other Immunogen Type corresponding to TRAP1
• Description: Mouse Monoclonal
• Gene: TRAP1
• Conjugate: Unconjugated

Product images

ab2721 staining TRAP1 in NCI-H460 cells by Immunocytochemistry/Immunofluorescence. Cells were grown on chamber slides and fixed with formaldehyde. Cells were probed without (right) or primary antibody (left) at a dilution of 1:200 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Green - TRAP1, Red (phalloidin) - F-actin, Blue (DAPI) - nuclei. Images were taken at 60X magnification.

Immunohistochemistry was performed on normal biopsies of deparaffinized Human liver tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and probed with a TRAP1 monoclonal antibody (ab2721) at a dilution of 1:20 or without primary antibody (negative control) overnight at 4�C in a humidified chamber. Tissues were washed 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.

Immunoprecipitation of TRAP1 using ab2721 visualized by Coomassie Blue staining.

ICC/IF image of ab2721 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2721, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

All lanes : Anti-TRAP1 antibody [TRAP1-6] (ab2721) at 1 µg/mlLane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

TRAP1 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Mouse monoclonal to TRAP1 (ab2721)and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). HepG2 whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2721. Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.Bands: 75kDa: TRAP1

Product References

Expression of TRAP1 predicts poor survival of malignant glioma patients. - Expression of TRAP1 predicts poor survival of malignant glioma patients.

Li S, Lv Q, Sun H, Xue Y, Wang P, Liu L, Li Z, Li Z, Tian X, Liu YH. J Mol Neurosci. 2015 Jan;55(1):62-8.

PINK1 drives Parkin self-association and HECT-like E3 activity upstream of - PINK1 drives Parkin self-association and HECT-like E3 activity upstream of

Lazarou M, Narendra DP, Jin SM, Tekle E, Banerjee S, Youle RJ. J Cell Biol. 2013 Jan 21;200(2):163-72.

Whole-exome resequencing reveals recessive mutations in TRAP1 in individuals with - Whole-exome resequencing reveals recessive mutations in TRAP1 in individuals with

Saisawat P, Kohl S, Hilger AC, Hwang DY, Yung Gee H, Dworschak GC, Tasic V, Pennimpede T, Natarajan S, Sperry E, Matassa DS, Stajic N, Bogdanovic R, de Blaauw I, Marcelis CL, Wijers CH, Bartels E, Schmiedeke E, Schmidt D, Marzheuser S, Grasshoff-Derr S, Holland-Cunz S, Ludwig M, Nothen MM, Draaken M, Brosens E, Heij H, Tibboel D, Herrmann BG, Solomon BD, de Klein A, van Rooij IA, Esposito F, Reutter HM, Hildebrandt F. Kidney Int. 2014 Jun;85(6):1310-7.

Broad activation of the ubiquitin-proteasome system by Parkin is critical for - Broad activation of the ubiquitin-proteasome system by Parkin is critical for

Chan NC, Salazar AM, Pham AH, Sweredoski MJ, Kolawa NJ, Graham RL, Hess S, Chan DC. Hum Mol Genet. 2011 May 1;20(9):1726-37.

Studies of phosphoproteomic changes induced by nucleophosmin-anaplastic lymphoma - Studies of phosphoproteomic changes induced by nucleophosmin-anaplastic lymphoma

Wu F, Wang P, Zhang J, Young LC, Lai R, Li L. Mol Cell Proteomics. 2010 Jul;9(7):1616-32.

Parkin is recruited selectively to impaired mitochondria and promotes their - Parkin is recruited selectively to impaired mitochondria and promotes their

Narendra D, Tanaka A, Suen DF, Youle RJ. J Cell Biol. 2008 Dec 1;183(5):795-803.

Hsp90 chaperone activity requires the full-length protein and interaction among - Hsp90 chaperone activity requires the full-length protein and interaction among

Johnson BD, Chadli A, Felts SJ, Bouhouche I, Catelli MG, Toft DO. J Biol Chem. 2000 Oct 20;275(42):32499-507.

The hsp90-related protein TRAP1 is a mitochondrial protein with distinct - The hsp90-related protein TRAP1 is a mitochondrial protein with distinct

Felts SJ, Owen BA, Nguyen P, Trepel J, Donner DB, Toft DO. J Biol Chem. 2000 Feb 4;275(5):3305-12.