RecombinantRabMAb

Recombinant Anti-MICA antibody [EPR24086-121] (ab259934)

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Flow Cytometry - Anti-MICA antibody [EPR24086-121] (ab259934)
  • Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
  • Immunoprecipitation - Anti-MICA antibody [EPR24086-121] (ab259934)
  • Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
  • Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
  • Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
  • Anti-MICA antibody [EPR24086-121] (ab259934)

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
  • Suitable for: IP, WB, Flow Cyt
  • Reacts with: Human

Overview

  • Product name

    Anti-MICA antibody [EPR24086-121]
    See all MICA primary antibodies

  • Description

    Rabbit monoclonal [EPR24086-121] to MICA

  • 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 within Human MICA aa 1-300. The exact sequence is proprietary.
    Database link: Q29983

  • 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

    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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR24086-121

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab259934 in the following tested applications.

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

Application Abreviews Notes
IP 1/30.
WB 1/1000. Detects a band of approximately 50, 60 kDa (predicted molecular weight: 43 kDa).
Flow Cyt 1/50.
  • Application notes
    Is unsuitable for ICC or IHC-P.

    • Target

    • Function

      Seems to have no role in antigen presentation. Acts as a stress-induced self-antigen that is recognized by gamma delta T-cells. Ligand for the KLRK1/NKG2D receptor. Binding to KLRK1 leads to cell lysis.

    • Tissue specificity

      Widely expressed with the exception of the central nervous system where it is absent. Expressed predominantly in gastric epithelium and also in monocytes, keratinocytes, endothelial cells, fibroblasts and in the outer layer of Hassal's corpuscles within the medulla of normal thymus. In skin, expressed mainly in the keratin layers, basal cells, ducts and follicles. Also expressed in many, but not all, epithelial tumors of lung, breast, kidney, ovary, prostate and colon. In thyomas, overexpressed in cortical and medullar epithelial cells. Tumors expressing MICA display increased levels of gamma delta T cells.

    • Involvement in disease

      Note=Anti-MICA antibodies and ligand shedding are involved in the progression of monoclonal gammopathy of undetermined significance (MGUS)to multiple myeloma.
      Genetic variations in MICA may be a cause of susceptibility to psoriasis type 1 (PSORS1) [MIM:177900]. Psoriasis is a common, chronic inflammatory disease of the skin with multifactorial etiology. It is characterized by red, scaly plaques usually found on the scalp, elbows and knees. These lesions are caused by abnormal keratinocyte proliferation and infiltration of inflammatory cells into the dermis and epidermis.
      Genetic variation in MICA is a cause of susceptibility to psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis).

    • Sequence similarities

      Belongs to the MHC class I family. MIC subfamily.
      Contains 1 Ig-like C1-type (immunoglobulin-like) domain.

    • Post-translational
      modifications

      N-glycosylated. Glycosylation is not essential for interaction with KLRK1/NKG2D but enhances complex formation.
      Proteolytically cleaved and released from the cell surface of tumor cells which impairs KLRK1/NKG2D expression and T-cell activation.

    • Cellular localization

      Cell membrane. Cytoplasm. Expressed on the cell surface in gastric epithelium, endothelial cells and fibroblasts and in the cytoplasm in keratinocytes and monocytes. Infection with human adenovirus 5 suppresses cell surface expression due to the adenoviral E3-19K protein which causes retention in the endoplasmic reticulum.
    • Information by UniProt

    • Database links

      • Alternative names

        • HLA class I antigen antibody
        • FLJ36918 antibody
        • FLJ60820 antibody
        • MGC111087 antibody
        • MGC21250 antibody
        • MHC class I chain related gene A protein antibody
        • MHC class I chain related protein A antibody
        • MHC class I chain related protein A HLA B HLA C antibody
        • MHC class I polypeptide related sequence A antibody
        • MHC class I polypeptide-related sequence A antibody
        • MHC class I related protein antibody
        • MIC A antibody
        • MIC-A antibody
        • micA antibody
        • MICA_HUMAN antibody
        • OTTHUMP00000029088 antibody
        • OTTHUMP00000044528 antibody
        • OTTHUMP00000165170 antibody
        • OTTHUMP00000165172 antibody
        • PERB11.1 antibody
        • Stress inducible class I homolog antibody
        see all

      Images

      • Flow Cytometry - Anti-MICA antibody [EPR24086-121] (ab259934)
        Flow Cytometry - Anti-MICA antibody [EPR24086-121] (ab259934)

        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] (ab259934)
        Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
        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?



        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] (ab259934)
        Immunoprecipitation - Anti-MICA antibody [EPR24086-121] (ab259934)

        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] (ab259934)
        Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
        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?



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

        Exposure time: 3 minutes.

         

      • Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
        Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
        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?



        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] (ab259934)
        Western blot - Anti-MICA antibody [EPR24086-121] (ab259934)
        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?



        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] (ab259934)
        Anti-MICA antibody [EPR24086-121] (ab259934)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (0)

      Publishing research using ab259934? Please let us know so that we can cite the reference in this datasheet.

      ab259934 has not yet been referenced specifically in any publications.

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