K562 human chronic myelogenous leukemia cell line was stained with Mouse Anti-Human MICA Monoclonal Antibody (Catalog # MAB1300, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.

MICA Specificity is Shown by Flow Cytometry in Knockout Cell Line.

MICA knockout K562 human myelogenous leukemia cell line was stained with Mouse Anti-Human MICA Monoclonal Antibody (Catalog # MAB1300, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram) followed by anti-Mouse IgG PE-conjugated secondary antibody (Catalog # F0102B). No staining in the MICA knockout K562 cell line was observed. View our protocol for Staining Membrane-associated Proteins.

Detection of Human MICA by Flow Cytometry

Multiple receptors and ligands are involved in NK cell-mediated lysis of activated CD4+ T cells.Role of (A) activating and (B) inhibitory NK receptors in NK cell degranulation. Left column: representative histograms (of n≥3) for surface expression of ligands on activated (thick black line) and resting CD4+ T cells (thin black line). Isotype-matched control Ig are represented by dashed line (activated CD4+ T) and filled histogram (resting CD4+ T). Middle- and right column: NK and CD4+ T cells were activated for 4 days in vitro as described, and co-cultured for 4 hours with 10 ug/mL mAb (or relevant isotype-matched control Ig). Degranulation is shown for CD56dim (middle column) and CD56bright (right column) NK cells. Representative histograms of surface expression of receptors on activated (thick black line) and resting NK cells (thin black line). Isotype-matched control Ig are represented by dashed line (activated NK) and filled histogram (resting NK). * P<0.05, ** P<0.005, *** P<0.001. (C) Sorted IL-2-activated CD56dim and CD56bright NK cells were co-cultured with 51Cr-labeled activated CD4+ T cells in a 51Cr-release assay with human IgG4 isotype control (•) or anti-NKG2A mAb (○). Data represents n = 3 experiments. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22384114), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human MICA Antibody

Application

Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.02-0.06 µg/mL of this antibody will block 50% of the binding of 50 ng/mL of Recombinant Human MICA Fc Chimera (Catalog # 1300-MA) to immobilized Recombinant Human NKG2D Fc Chimera (Catalog # 1299-NK) coated at 2 µg/mL (100 µL/well). At 0.4 μg/mL, this antibody will block >90% of the binding.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: K562 human chronic myelogenous leukemia cell line

Knockout Validated

0.25 µg/10⁶ cells
Sample: MICA is specifically detected in K562 myelogenous leukemia parental cell line but is not detectable in MICA knockout K562 cell line.

Western Blot

1 µg/mL
Sample: Recombinant Human MICA Fc Chimera (Catalog # 1300-MA)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: MICA

MICA (MHC class I chain-related gene A) is a transmembrane glycoprotein that functions as a ligand for human NKG2D. A closely related protein, MICB, shares 85% amino acid identity with MICA. These proteins are distantly related to the MHC class I proteins. They possess three extracellular Ig-like domains, but they have no capacity to bind peptide or interact with beta2-microglobulin. The genes encoding these proteins are found within the Major Histocompatibility Complex on human chromosome 6. The MICA locus is highly polymorphic with more than 50 recognized human alleles. MICA is absent from most cells but is frequently expressed in epithelial tumors and can be induced by bacterial and viral infections. MICA is a ligand for human NKG2D, an activating receptor expressed on NK cells, NKT cells, gamma delta T cells, and CD8⁺ alpha beta T cells. Recognition of MICA by NKG2D results in the activation of cytolytic activity and/or cytokine production by these effector cells. MICA recognition is involved in tumor surveillance, viral infections, and autoimmune diseases.

References

Groh, V. et al. (2001) Nature Immunol. 2:255.
Stephens, H. (2001) Trends Immunol. 22:378.
Bauer, S. et al. (1999) Science 285:727.
Groh, V. et al. (2002) Nature 419:734.
Steinle, A. et al. (2001) Immunogenetics 53:279.
Pende, D. et al. (2002) Cancer Res. 62:6178.
NKG2D and its Ligands (2002) http://www.RnDSystems.com

Long Name

MHC Class I-related Protein A

Alternate Names

FLJ60820, MGC111087, PERB11.1

Entrez Gene IDs

100507436 (Human)

Gene Symbol

MICA

Additional MICA Products

Product Documents for Human MICA Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human MICA Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human MICA Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB1300

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

View all MICA Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Endotoxin Level

Scientific Data Images for Human MICA Antibody

Detection of MICA in K562 Human Cell Line by Flow Cytometry.