Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24086-121] to MICA - BSA and Azide free
  • Suitable for: IP, WB, Flow Cyt
  • Reacts with: Human

Overview

  • Product name

    Anti-MICA antibody [EPR24086-121] - BSA and Azide free
    See all MICA primary antibodies

  • Description

    Rabbit monoclonal [EPR24086-121] to MICA - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, Flow Cytmore details
    Unsuitable for: ICC or IHC-P

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa and HUVEC whole cell lysates; Human breast cancer and colon cancer tissue lysates; His-tagged human recombinant protein MHC class I polypeptide-related sequence A. Flow cyt: HeLa cells. IP: HUVEC whole cell lysate.

  • General notes

    ab276141 is the carrier-free version of ab259934. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab276141 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Flow Cytometry - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    Flow Cytometry - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)

    This data was developed using ab259934, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of THP-1 (Human monocytic leukemia monocyte, left)/ HeLa (human cervix adenocarcinoma epithelial cell, right) cells labelling MICA with ab259934 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730)  isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: THP-1 (PMID: 28154561).

    Gated on viable cells.

  • Western blot - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    Western blot - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    All lanes : Anti-MICA antibody [EPR24086-121] (ab259934) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : THP-1 (Human monocytic leukemia monocyte) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab259934, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Negative control: THP-1 (PMID: 28154561).

    MICA is a glycoprotein and can be de-glycosylated by PNGase F. The molecular mass observed is consistent with the literature (PMID: 10359807, 9396860).

    Exposure time: 3 minutes.

  • Immunoprecipitation - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    Immunoprecipitation - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)

    This data was developed using ab259934, the same antibody clone in a different buffer formulation.

    MICA was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate with ab259934 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259934 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HUVEC (human umbilical vein endothelial cell) whole cell lysate 10 ug

    Lane 2: ab259934 IP in HUVEC whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259934 in HUVEC whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

    This blot was developed using a higher sensitivity ECL substrate.

  • Western blot - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    Western blot - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    All lanes : Anti-MICA antibody [EPR24086-121] (ab259934) at 1/1000 dilution

    Lane 1 : His-tagged human recombinant protein MHC class I polypeptide-related sequence A, 10 ng
    Lane 2 : His-tagged human recombinant protein MHC class I polypeptide-related sequence B, 10 ng

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?



    This data was developed using ab259934, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Western blot - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    Western blot - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    All lanes : Anti-MICA antibody [EPR24086-121] (ab259934) at 1/1000 dilution

    Lane 1 : Human breast cancer tissue lysate
    Lane 2 : Human colon cancer tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

    Predicted band size: 43 kDa
    Observed band size: 50-60 kDa why is the actual band size different from the predicted?



    This data was developed using ab259934, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    MICA is a glycoprotein and can be de-glycosylated by PNGase F. The molecular mass observed is consistent with the literature (PMID: 10359807, 9396860).

    Exposure time: 3 minutes.

  • Western blot - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    Western blot - Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    All lanes : Anti-MICA antibody [EPR24086-121] (ab259934) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa whole cell lysate treated with PNGase F
    Lane 3 : HUVEC (human umbilical vein endothelial cell) whole cell lysate
    Lane 4 : HUVEC whole cell lysate treated with PNGase F

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa
    Observed band size: 32,50-60 kDa why is the actual band size different from the predicted?



    This data was developed using ab259934, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    MICA is a glycoprotein and can be de-glycosylated by PNGase F. The molecular mass observed is consistent with the literature (PMID: 10359807, 9396860).

    Exposure time: 3 minutes.

  • Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)
    Anti-MICA antibody [EPR24086-121] - BSA and Azide free (ab276141)

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