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MICA Recombinant Rabbit Monoclonal Antibody [PSH13-60] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751479

MICA Recombinant Rabbit Monoclonal Antibody [PSH13-60] - BSA and Azide free

Application

  • WB

  • FC

  • IP

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

MICA Recombinant Rabbit Monoclonal Antibody [PSH13-60] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human MICA aa 1-328.

Species Reactivity

Human

Validated Applications

WB, FC, IP

Molecular Weight

Predicted band size: 43 kDa

Positive Control

HeLa cell lysate, MCF7 cell lysate, OVCAR-3 cell lysate, HeLa.

Conjugation

unconjugated

Clone Number

PSH13-60

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • FC

  • 1:5,000

  • IP

  • 1-2μg/sample

Target

Function

MHC class I polypeptide–related sequence A (MICA) is a highly polymorphic cell surface glycoprotein encoded by the MICA gene located within MHC locus. MICA is related to MHC class I and it has similar domain structure, however, it is not associated with β2-microglobulin nor binds peptides as conventional MHC class I molecules do. MICA rather functions as a stress-induced ligand (as a danger signal) for integral membrane protein receptor NKG2D ("natural-killer group 2, member D"). MICA is broadly recognized by NK cells, γδ T cells, and CD8+ αβ T cells which carry NKG2D receptor on their cell surface and which are activated via this interaction.

Background References

1. Li XC. MICA is a histocompatibility antigen in transplantation. Am J Transplant. 2022 Sep

2. Allison SJ. MICA in kidney transplants. Nat Rev Nephrol. 2022 May

Subcellular Location

Cell membrane, Cytoplasm.

UNIPROT

SwissProt: Q29983 Human

Synonyms

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

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of MICA on different lysates with Rabbit anti-MICA antibody (<a href="/products/HA751479" style="font-weight: bold;text-decoration: underline;">HA751479</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate (10 µg/Lane)<br />Lane 2: MCF7 cell lysate (25 µg/Lane)<br />Lane 3: THP-1 cell lysate (negative) (25 µg/Lane)<br />Lane 4: OVCAR-3 cell lysate (25 µg/Lane)<br /><br />Predicted band size: 43 kDa<br />Observed band size: 60 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751479" style="font-weight: bold;text-decoration: underline;">HA751479</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of MICA on different lysates with Rabbit anti-MICA antibody (HA751479) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate (10 µg/Lane)
    Lane 2: MCF7 cell lysate (25 µg/Lane)
    Lane 3: THP-1 cell lysate (negative) (25 µg/Lane)
    Lane 4: OVCAR-3 cell lysate (25 µg/Lane)

    Predicted band size: 43 kDa
    Observed band size: 60 kDa

    Exposure time: 20 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751479) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Flow cytometric analysis of THP-1 (left, negative) and HeLa (right, positive) cells labeling MICA.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751479" style="font-weight: bold;text-decoration: underline;">HA751479</a>, 1/5,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    ☑ Relative expression (RE)

    Flow cytometric analysis of THP-1 (left, negative) and HeLa (right, positive) cells labeling MICA.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751479, 1/5,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • MICA was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA751479" style="font-weight: bold;text-decoration: underline;">HA751479</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751479" style="font-weight: bold;text-decoration: underline;">HA751479</a> at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA751479" style="font-weight: bold;text-decoration: underline;">HA751479</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751479" style="font-weight: bold;text-decoration: underline;">HA751479</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>)<br />Exposure time: 59 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    MICA was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751479 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751479 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA751479 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA751479 in HeLa cell lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 59 seconds; ECL: K1801

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"