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Signal Transduction Protein Trafficking Chaperones Heat Shock Proteins
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Anti-TRAP1 antibody [TRAP1-6] (ab2721)

  • Datasheet
  • SDS
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Immunocytochemistry/ Immunofluorescence - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
  • Immunoprecipitation - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
  • Immunocytochemistry/ Immunofluorescence - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
  • Western blot - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
  • Immunoprecipitation - 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

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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 & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Other Immunogen Type corresponding to TRAP1. Purified recombinant TRAP1.

  • Positive control

    • ICC: 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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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

    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Heat Shock Proteins
    • Cell Biology
    • Apoptosis
    • Mitochondrial
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Metabolism
    • Types of disease
    • Cancer
    • Cancer
    • Cell Death
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Recombinant Protein

    • Recombinant human TRAP1 protein (ab123775)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Human
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.
  • Target information above from: UniProt accession Q12931 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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
      • HSP75 antibody
      • HSP90L antibody
      • mitochondrial antibody
      • TNF receptor associated protein 1 antibody
      • TNFR-associated protein 1 antibody
      • TRAP-1 antibody
      • Trap1 antibody
      • TRAP1_HUMAN antibody
      • Tumor necrosis factor type 1 receptor-associated protein antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      Immunocytochemistry/ Immunofluorescence - Anti-TRAP1 antibody [TRAP1-6] (ab2721)

      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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAP1 antibody [TRAP1-6] (ab2721)

      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 - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      Immunoprecipitation - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      Immunoprecipitation of TRAP1 using ab2721 visualized by Coomassie Blue staining.
    • Immunocytochemistry/ Immunofluorescence - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      Immunocytochemistry/ Immunofluorescence - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      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.

    • Western blot - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      Western blot - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      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.
    • Immunoprecipitation - Anti-TRAP1 antibody [TRAP1-6] (ab2721)
      Immunoprecipitation - Anti-TRAP1 antibody [TRAP1-6] (ab2721)

      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

    Protocols

    • Recommended ICC protocol with ab2721

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (13)

    Publishing research using ab2721? Please let us know so that we can cite the reference in this datasheet.

    ab2721 has been referenced in 13 publications.

    • 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
    View all Publications for this product

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