Anti-TRAP1 antibody [TRAP1-6]

• 产品名称

  Anti-TRAP1抗体[TRAP1-6]
  参阅全部 TRAP1 一抗

• 描述

  小鼠单克隆抗体[TRAP1-6] to TRAP1

• 宿主

  Mouse

• 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
  

• 免疫原

  Recombinant full length protein corresponding to Human TRAP1. Purified recombinant TRAP1.
  Database link: Q12931

• 阳性对照

• ICC/IF: PC-3-M cells

• 常规说明

  
  

  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

• 形式

  Liquid

• 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

• 纯度

  Affinity purified

• 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.

• 克隆

  单克隆

• 克隆编号

  TRAP1-6

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab2721于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

• Entrez Gene: 10131 Human

• Entrez Gene: 68015 Mouse

• Omim: 606219 Human

• SwissProt: Q12931 Human

• SwissProt: Q9CQN1 Mouse

• Unigene: 30345 Human

• Unigene: 123366 Mouse

• 别名

• 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

• Heat shock protein 75 kDa antibody

• Heat shock protein 75 kDa, mitochondrial antibody

• HSP 75 antibody

• 

  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.

• 

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

  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 (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 of TRAP1 using ab2721 visualized by Coomassie Blue staining.

• 

  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)

  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 kDawhy 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)

  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