Anti-TRAP1 antibody [TRAP1-6] (ab2721)
Key features and details
- Mouse monoclonal [TRAP1-6] to TRAP1
- Suitable for: IHC-P, IP, WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG1
Overview
-
Product name
Anti-TRAP1 antibody [TRAP1-6]
See all TRAP1 primary antibodies -
Description
Mouse monoclonal [TRAP1-6] to TRAP1 -
Host species
Mouse -
Specificity
Detects tumor necrosis factor receptor-associated protein (TRAP1) from human tissues. -
Tested applications
Suitable for: IHC-P, IP, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant full length protein corresponding to Human TRAP1. Purified recombinant TRAP1.
Database link: Q12931 -
Positive control
- ICC/IF: PC-3-M cells
-
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
-
Form
Liquid -
Storage instructions
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. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading...
-
Purity
Affinity purified -
Primary antibody notes
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. -
Clonality
Monoclonal -
Clone number
TRAP1-6 -
Isotype
IgG1 -
Research areas
Associated products
-
Compatible Secondaries
-
Isotype control
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
---|---|---|
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).
|
|
ICC/IF |
1/250.
|
Notes |
---|
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). |
ICC/IF
1/250. |
Target
-
Function
Chaperone that expresses an ATPase activity. -
Tissue specificity
Found in skeletal muscle, liver, heart, brain, kidney, pancreas, lung and placenta. -
Sequence similarities
Belongs to the heat shock protein 90 family. -
Cellular localization
Mitochondrion. - Information by UniProt
-
Database links
- Entrez Gene: 10131 Human
- Entrez Gene: 68015 Mouse
- Omim: 606219 Human
- SwissProt: Q12931 Human
- SwissProt: Q9CQN1 Mouse
- Unigene: 30345 Human
- Unigene: 123366 Mouse
-
Alternative names
- Heat shock protein 75 kDa antibody
- Heat shock protein 75 kDa, mitochondrial antibody
- HSP 75 antibody
see all
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.
-
All lanes : Anti-TRAP1 antibody [TRAP1-6] (ab2721) at 1/2000 dilution
Lane 1 : HeLa whole cell lysate
Lane 2 : Hep G2 whole cell lysate
Lane 3 : HEK-293 whole cell lysate
Lane 4 : K-562 whole cell lysate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/40000 dilution
Predicted band size: 80 kDa -
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.
Secondary
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
Datasheets and documents
-
SDS download
-
Datasheet download
References (14)
ab2721 has been referenced in 14 publications.
- Xue Y et al. Interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) accelerates osteoclast formation by regulating signal transducer and activator of transcription 3 (STAT3) signalling. Bioengineered 13:2285-2295 (2022). PubMed: 35034537
- 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