Key features and details
- Mouse monoclonal [TRAP1-6] to TRAP1
- Suitable for: IHC-P, IP, WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG1
参阅全部 TRAP1 一抗
描述小鼠单克隆抗体[TRAP1-6] to TRAP1
特异性Detects tumor necrosis factor receptor-associated protein (TRAP1) from human tissues.
经测试应用适用于: IHC-P, IP, WB, ICC/IFmore details
种属反应性与反应: Mouse, Human
Other Immunogen Type corresponding to TRAP1. Purified recombinant TRAP1.
- ICC: PC-3-M cells
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
存储溶液Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS
Concentration information loading...
Primary antibody说明Immunofluorescence staining of TRAP1 in PC-3-M cells with this antibody produces a pattern consistent with mitochondrial staining. Immunoprecipitation of TRAP1 using this antibody fails to co-precipitate p23, Hop, or CyP40 suggesting TRAP1’s inability to associate with these co-chaperones.
Our Abpromise guarantee covers the use of ab2721 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/10 - 1/100.|
|IP||Use at an assay dependent concentration.|
|WB||1/2000. Detects a band of approximately 76 kDa (predicted molecular weight: 80 kDa).|
功能Chaperone that expresses an ATPase activity.
组织特异性Found in skeletal muscle, liver, heart, brain, kidney, pancreas, lung and placenta.
序列相似性Belongs to the heat shock protein 90 family.
- Information by UniProt
- Heat shock protein 75 kDa antibody
- Heat shock protein 75 kDa, mitochondrial antibody
- HSP 75 antibody
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/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 80 kDa
Observed band size: 76 kDa why is the actual band size different from the predicted?
Additional bands at: 40 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 16 minutes
The band observed at 76 kDa could potentially be a cleaved form of TRAP1 due to the presence of a 59 amino acid transit peptide.
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
ab2721 被引用在 13 文献中.
- MacDonald JA et al. A nanoscale, multi-parametric flow cytometry-based platform to study mitochondrial heterogeneity and mitochondrial DNA dynamics. Commun Biol 2:258 (2019). PubMed: 31312727
- Xiao B et al. Reactive oxygen species trigger Parkin/PINK1 pathway-dependent mitophagy by inducing mitochondrial recruitment of Parkin. J Biol Chem 292:16697-16708 (2017). PubMed: 28848050
- Roundhill E et al. Localization of MRP-1 to the outer mitochondrial membrane by the chaperone protein HSP90ß. FASEB J 30:1712-23 (2016). PubMed: 26722004
- Du Y et al. Wnt3a is critical for endothelial progenitor cell-mediated neural stem cell proliferation and differentiation. Mol Med Rep 14:2473-82 (2016). WB . PubMed: 27484039
- McLelland GL et al. Parkin and PINK1 function in a vesicular trafficking pathway regulating mitochondrial quality control. EMBO J 33:282-95 (2014). PubMed: 24446486