Recombinant
RabMAb

Recombinant Anti-MICA + MICB antibody [EPR22031] (ab224702)

Overview

  • Product name

    Anti-MICA + MICB antibody [EPR22031]
    See all MICA + MICB primary antibodies
  • Description

    Rabbit monoclonal [EPR22031] to MICA + MICB
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human MICA + MICB aa 1-400. The exact sequence is proprietary.
    Database link: Q29983

  • Positive control

    • WB: His-tagged MICB recombinant protein expressed by the E. coli system (aa23-383 with 310-330 replaced by GGGS). His-tagged MICA recombinant protein expressed by the E. coli system (aa24-383 with 308-328 replaced by GGGGS). PNGase F treated HeLa and K562 cell lysate. U266B1, HUVEC, K562 and HeLa whole cell lysate. IHC-P: Human ovary cancer, lung cancer and stomach tissue.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

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: PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22031
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab224702 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
WB 1/1000. Detects a band of approximately 50-60 kDa (predicted molecular weight: 42 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    Anti-MICA antibodies and ligand shedding are involved in the progression of monoclonal gammopathy of undetermined significance (MGUS)to multiple myeloma.
    Psoriasis 1
    Psoriatic arthritis
  • 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

    • MHC class I chain-related protein A antibody
    • MHC class I chain-related protein B antibody
    • MHC class I polypeptide related sequence A antibody
    • MHC class I polypeptide related sequence B antibody
    • MHC class I polypeptide-related sequence A antibody
    • MIC-A antibody
    • micA antibody
    • MICA_HUMAN antibody
    • MICB antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling MICA with ab224702 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on epithelium of human stomach (PMID: 28334733) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-MICA + MICB antibody [EPR22031] (ab224702) at 1/1000 dilution

    Lane 1 : K562 (human chronic myelogenous leukemia cell line from bone marrow), whole cell lysate
    Lane 2 : K562 whole cell lysate treated with PNGase F
    Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
    Lane 4 : HeLa whole cell lysate treated with PNGase F
    Lane 5 : HUVEC (human umbilical vein endothelial cell line), whole cell lysate
    Lane 6 : U266B1 (human multiple myeloma B lymphocyte cell line), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 42 kDa
    Observed band size: 40,50-60 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Bands between 50-60kDa represent the glycosylated form. After PNGase F treatment, the de-glycosylated form appears around 40kDa.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 10359807; PMID: 9396860).

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue labeling MICA with ab224702 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on human ovary cancer (PMID: 28334733) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling MICA with ab224702 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on human lung cancer (PMID: 28334733) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-MICA + MICB antibody [EPR22031] (ab224702) at 1/5000 dilution

    Lane 1 : His-tagged MICB recombinant protein expressed in E.coli system (aa23-383 with 310-330 replaced by GGGS), 10 ng
    Lane 2 : His-tagged MICA recombinant protein expressed in E.coli system (aa24-383 with 308-328 replaced by GGGGS) 10 ng

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 42 kDa


    Exposure time: 15 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

References

ab224702 has not yet been referenced specifically in any publications.

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