Anti-TOMM20抗体- Mitochondrial Marker (ab78547)

概述

  • 产品名称Anti-TOMM20抗体- Mitochondrial Marker
    参阅全部 TOMM20 一抗
  • 描述
    兔多克隆抗体to TOMM20 - Mitochondrial Marker
  • 经测试应用适用于: ICC/IF, IP, WB, IHC-Pmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse, Rat, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Human TOMM20.

    (Peptide available as ab88207.)

  • 阳性对照
    • This antibody gave a positive signal in HepG2 whole cell lysate.

性能

应用

Our Abpromise guarantee covers the use of ab78547 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF Use a concentration of 1 µg/ml.
IP Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

Anti-TOMM20 antibody - Mitochondrial Marker 图像

  • ICC/IF image of ab78547 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 (ab78547, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293, MCF-7 cells at 1µg/ml, and in 100% Methanol fixed (5 min) HepG2, HeLa, Hek293, MCF-7 cells at 1µg/ml.
  • Anti-TOMM20 antibody - Mitochondrial Marker (ab78547) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 16 kDa
    Observed band size : 16 kDa
    Additional bands at : 70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
  • IHC image of TOMM20 staining in Human Liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78547, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • TOMM20 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to TOMM20 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 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 ab78547.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 16kDa; TOMM20.

Anti-TOMM20 antibody - Mitochondrial Marker (ab78547)参考文献

This product has been referenced in:
  • Kim SJ  et al. Hepatitis C virus induces the mitochondrial translocation of Parkin and subsequent mitophagy. PLoS Pathog 9:e1003285 (2013). Human . Read more (PubMed: 23555273) »
  • Kim SJ  et al. Hepatitis B virus disrupts mitochondrial dynamics: induces fission and mitophagy to attenuate apoptosis. PLoS Pathog 9:e1003722 (2013). Human . Read more (PubMed: 24339771) »

See all 3 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (mouse embryonic fibroblast)
Specification mouse embryonic fibroblast
Fixative Formaldehyde
Permeabilization Yes - 0.5% Triton X
Blocking step BSA and image-iTFX (invitrogen) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Abcam user community

Verified customer

提交于 Feb 15 2013

Thank you for your response.

My colleague is out of office this week and she has asked me to look after her customers.

There seems to be a faint band at 15 kDa and another stronger one at 60 kDa in the Western blot image you have se...

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Thank you for your response.

My colleague Kate is out of office this week and she has asked me to look after her colleagues.

I understand that this vial was a free of charge replacement. Could you please summarize if the problem wit...

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Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this second antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100/PBS
Blocking step 0.5% BSA, 0.2% fish skin gelatin/PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 28 2010

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Xenopus laevis Cell lysate - whole cell (meiotic egg extract)
Loading amount 25 µg
Specification meiotic egg extract
Gel Running Conditions Reduced Denaturing (15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 21 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"