Anti-PGRMC1抗体(ab80941)

概述

  • 产品名称Anti-PGRMC1抗体
    参阅全部 PGRMC1 一抗
  • 描述
    兔多克隆抗体to PGRMC1
  • 经测试应用适用于: ICC/IF, WB, IHC-Pmore details
  • 种属反应性
    与反应: Human
    预测可用于: Cow, Pig, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human PGRMC1.

    (Peptide available as ab88945.)

  • 阳性对照
    • This antibody gave a positive signal in Human Liver and Ovary tissue lysates and the following whole cell lysates; T47D, MCF7, HEK293, and HepG2. This antibody gave a positive result in IHC in the following FFPE tissue: Human liver cancer.

性能

应用

Our Abpromise guarantee covers the use of ab80941 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.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 21 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

靶标

Anti-PGRMC1 antibody 图像

  • IHC image of PGRMC1 staining in Human liver cancer formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab80941, 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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-PGRMC1 antibody (ab80941) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : T47D whole cell lysate (ab14899)
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : Human ovary tissue lysate - total protein (ab30222)

    Lysates/proteins at 10 µg per lane.

    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 : 21 kDa
    Observed band size : 25 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa,150 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute
    Membrane-associated progesterone receptor component 1 (PGRMC1) contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight (25 kDa) than predicted (21 kDa).
  • ICC/IF image of ab80941 stained HeLa 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 (ab80941, 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.

Anti-PGRMC1 antibody (ab80941)参考文献

ab80941 has not yet been referenced specifically in any publications.

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Fibroblast)
Specification Fibroblast
Fixative Paraformaldehyde
Permeabilization Yes - 1% Triton X-100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 25°C
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提交于 Apr 15 2013

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