K562 human chronic myelogenous leukemia cell line was stained with Mouse Anti-Human MICB Monoclonal Antibody (Catalog # MAB1599, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B).

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

MICB knockout K562 human myelogenous leukemia cell line was stained with Mouse Anti-Human MICB Monoclonal Antibody (Catalog # MAB1599, 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 MICB knockout K562 cell line was observed. View our protocol for Staining Membrane-associated Proteins.

Detection of Human MICB 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.

Detection of Mouse MICB by Flow Cytometry

Hydroxycitrate reduces MICA expression in activated T cells and multiple cancer cells. (A) MICA mRNA analyzed by quantitative RT-PCR in total RNA purified from HEK293 cells after more than 30 passages in glucose (Glc) and galactose (Gal). MICA expression is normalized to housekeeping gene RPLP0 and displayed as mean ± SEM from six independent experiments. (B) MICA surface expression analyzed by flow cytometry of Glc and Gal cells at basal levels. Dot plots are representative of at least three independent experiments. Grid is set to ∼5% of corresponding isotype control stainings. (C) Mitochondrial stress test on HEK293 cultivated in Glc or Gal under same conditions as in Figure 4A. The graph is baselined to measuring point three and displays mean ± SEM from two independent experiments. (D) MICA/B surface expression of peripheral blood lymphocytes (PBLs) activated for 3 days in Glc or Gal growth medium prior to 18 h treatment with FR901228 (20 ng/mL). Grids in dot plots are set to ∼5% of corresponding isotype control staining and dot plots are representative of seven different donors. The bar graph displays mean ± SEM of isotype-corrected MICA/B MFI ( deltaMFI) from seven donors. Left panel is zoomed in on the difference between untreated Glc and Gal PBLs. (E,F) HEK293 MGAT5 knockout (KO) cells were treated with (E) 2DG (20 mM) or (F) hydroxycitrate (HC) (15 mM) in addition to PBS (UT), citrate (10 mM), or GlcNAc (25 mM) for 22–24 h. Bar graphs display MICA surface expression as mean ± SEM of deltaMFI values from three independent experiments. Data of UT samples share values with UT samples in Figure 3H. (G) MICA surface expression in several cancer cell lines after 18 or 42 h treatment with HC (10 mM). deltaMFI values are normalized to UT control and shown as mean ± SEM from at least three independent experiments. (H,I) MICA surface expression (H) and NKG2D-fc binding (I) in cancer cell lines after 2.5 h treatment with HC (10 mM) prior to 18 h stimulation with FR901228 (FR, 20 ng/mL) or sodium butyrate (But, 5 mM). Bar graphs display MICA surface expression as mean ± SEM of deltaMFI values (H), or NKG2D-fc surface binding as ± SEM of deltaMFI normalized to untreated (UT) control (I), from three independent experiments. Statistical analysis was performed by unpaired t-test with Welch’s correction in (A,E,F), ratio paired t-test in (D), one-sample t-test in (G), and two-way ANOVA with Bonferroni’s multiple comparison test (H,I). *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32849657), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human MICB Antibody

Application

Recommended Usage

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

MICB is specifically detected in K562 myelogenous leukemia parental cell line but is not detectable in MICB knockout K562 cell line.

Western Blot

1 µg/mL
Sample: Recombinant Human MICB Fc Chimera, aa 23-298 (Catalog # 1599-MB)

Human MICB Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)

Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:

Detection Reagent: Human MICB Biotinylated Antibody (Catalog # BAF1599)
Standard: Recombinant Human MICB (aa 23-298) Fc Chimera Protein, CF (Catalog # 1599-MB)

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

Reviewed Applications

Read 1 review rated 5 using MAB1599 in the following applications:

Flow Cytometry (1 Review)

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: MICB

MICB (MHC class I chain-related gene B) is a transmembrane glycoprotein that functions as a ligand for NKG2D. A closely related protein, MICA, shares 85% amino acid identity with MICB. These 2 proteins are distantly related to the MHC class I proteins. MICA and MICB (MICA/B) possess three extracellular immunoglobulin-like domains, but have no capacity to bind peptide or interact with beta2-microglobulin. The genes encoding MICA/B are found within the major histocompatibility complex on human chromosome 6. The MICB locus is polymorphic with more than 15 recognized human alleles. MICA/B are minimally expressed on normal cells, but are frequently expressed on epithelial tumors and can be induced by bacterial and viral infections. MICA/B are ligands for NKG2D, an activating receptor expressed on NK cells, NKT cells, gamma delta T cells, and CD8⁺ alpha beta T cells. Recognition of MICA/B by NKG2D results in the activation of cytolytic activity and/or cytokine production by these effector cells. MICA/B recognition is involved in tumor surveillance, viral infections, and autoimmune diseases. The release of soluble forms of MICA/B from tumors down-regulates NKG2D surface expression on effector cells resulting in the impairment of anti-tumor immune response (1-7).

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.
Salih, H. et al. (2003) Blood 102:1389.

Long Name

MHC Class I-related Protein B

Alternate Names

MHC class I chain-related protein B, MHC class I mic-B antigen, MHC class I polypeptide-related sequence B, MIC-B, PERB11.2MHC class I-like molecule PERB11.2-IMX, stress inducible class I homolog

Entrez Gene IDs

4277 (Human)

Gene Symbol

MICB

UniProt

CAI18747

Additional MICB Products

Product Documents for Human MICB Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human MICB Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human MICB Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB1599

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications