PRODUCT CODE: ER1803-95

MICA + MICB Rabbit Polyclonal Antibody (ER1803-95)

Applications

  • WB

  • ICC

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of MICA + MICB on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control:<br />
Lane 1: U937<br />
Lane 2: Mouse bone marrow
  • Western blot analysis of MICA + MICB on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control:<br />
Lane 1: U937<br />
Lane 2: Mouse bone marrow
  • ICC staining MICA + MICB in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with MICA + MICB at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat skin tissue using anti-MICA + MICB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (ER1803-95) at 1/50 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using anti-MICA + MICB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (ER1803-95) at 1/50 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-MICA + MICB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (ER1803-95) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-MICA + MICB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (ER1803-95) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue using anti-MICA + MICB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (ER1803-95) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
Western blot analysis of MICA + MICB on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: U937
Lane 2: Mouse bone marrow

Applications

  • WB

  • ICC

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

MICA + MICB Rabbit Polyclonal Antibody (ER1803-95)

Immunogen

Recombinant protein within human mica + micb aa 180-310.

Host

Rabbit

Positive Control

U937, Mouse bone marrow, A431, rat skin tissue, human lung cancer tissue, human colon tissue, human breast carcinoma tissue, mouse stomach tissue.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein affinity purified.

MOLECULAR WEIGHT

42 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB:1:500-1:2,000

  • ICC:1:50-1:100

  • IHC-P:1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

MICA + MICB

SYNONYMS

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

SEQUENCE SIMILARITIES

Belongs to the MHC class I family. MIC subfamily.

TISSUE SPECIFICITY

Widely expressed with the exception of the central nervous system where it is absent. Expressed in many, but not all, epithelial tumors of lung, breast, kidney, ovary, prostate and colon. In hepatocellular carcinomas, expressed in tumor cells but not in surrounding non-cancerous tissue.

POST-TRANSLATIONAL MODIFICATION

Proteolytically cleaved and released from the cell surface of tumor cells.

SUBCELLULAR LOCATION

Cell membrane. Cytoplasm.

FUNCTION

MHC class I polypeptide-related sequence A/B (MICA/B) is a cell surface glycoprotein encoded by the MICA/B gene located within MHC locus. MICA and MICB are related to MHC class I and has similar domain structure, which is made up of external α1α2α3 domain, transmembrane segment and C-terminal cytoplasmic tail. MICA/B rather functions as a stress-induced ligand for NKG2D receptor. MICA is broadly recognized by NK cells, γδ T cells, and CD8+ αβ T cells which carry NKG2D receptor on their cell surface. As a result of NKG2D-MICA engagement, effector cytolytic responses of T cells and NK cells against epithelial tumor cells expressing MICA are initiated. MICA/B are not associated with β2-microglobulin nor binds peptides as conventional MHC class I molecules do. The heat shock stress pathway is involved in the regulation of MICA expression as transcription of MICA is regulated by promoter heat shock element.