ANTI-MOUSE MHCII/I-A (NIMR-4) COLORWHEEL DYE-READY MAB

General description

ColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel^® antibody with any ColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel^® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel^® technology requires both ColorWheel^® antibody and ColorWheel^® dye for Flow Cytometry application.

We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This product is Preservative-free, lyophilized product for enhanced stability and allow for ambient shipping and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.

Specificity

Clone NIRM-4 (NIM-R4) is a rat monoclonal antibody that detects murine H-2 class II histocompatibility antigen, A-B alpha chain (MHCII/I-A).

Immunogen

Murine B-lymphocytes.

Application

Quality Control Testing

Evaluated by Flow Cytometry in Mouse splenocytes.

Flow Cytometry Analysis: Staining of one million C57BL/6 mouse splenocytes was performed with APC-conjugated anti-mouse CD3e and with 5 μL of a 1:1 mixture of Cat. No. CWA-1135, Anti-Mouse MHCII/I-A (NIMR-4) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye (right figure) or an equivalent amount of PE-conjugated Rat IgG2b isotype control. MHCII/I-A expressing cells were observed in CD3e- population.

Tested Applications

Flow Cytometry Analysis: Staining of one million C57BL/6 mouse splenocytes was performed with PE anti-mouse CD3e and with 5 μL of a 1:1 mixture of Cat. No. CWA-1135, Anti-Mouse MHCII/I-A (NIMR-4) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheelColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or an equivalent amount of APC-conjugated Rat IgG2b isotype control. MHCII/I-A expressing cells were observed in CD3e- population.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Evaluated by Flow Cytometry in Mouse splenocytes.

Flow Cytometry Analysis: Staining of one million C57BL/6 mouse splenocytes was performed with APC-conjugated anti-mouse CD3e and with 5 μL of a 1:1 mixture of Cat. No. CWA-1135, Anti-Mouse MHCII/I-A (NIMR-4) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye (right figure) or an equivalent amount of PE-conjugated Rat IgG2b isotype control. MHCII/I-A expressing cells were observed in CD3e- population.

Compatibility

Requires ColorWheel^® Dye (Sold Separately)

Target description

H-2 class II histocompatibility antigen, A-B alpha chain (UniProt: P14434; also known as IAalpha) is encoded by the H2Aa gene (Gene ID: 14960) in murine species. Major Histocompatibility Complex (MHC) class II is a heterodimeric transmembrane protein that is expressed on antigen-presenting cells including dendritic cells, macrophages, and B cells. It contains and a β subunits that are assembled into heterodimers in the endoplasmic reticulum, where they are complexed to an invariant chain (CD74), a dedicated chaperone protein that directs MHC class II into a low-pH, late-stage endosomal compartment, known as the MHC II compartment (MIIC). Within MIIC, proteases cleave invariant chain and leave a nested set of invariant chain fragments known as class II invariant chain-associated peptides (CLIP), which temporarily occupies the peptide binding groove of the MHC class II molecule. A catalytic protein, HLA-DM, exchanges CLIP for peptides that bind MHC class II with high-affinity. In addition to presenting peptide antigens to T cells, MHC class II can also initiate intracellular signaling pathways in antigen-presenting cells that modulate their proliferation, maturation, and apoptosis. MHC class II (1-A) is a single pass type I membrane protein that located in external side of plasma membrane and lysosome. It is synthesized with a signal peptide (aa 1-23), which is subsequently cleaved off to produce the mature form that contains an extracellular domain (aa 24-218), a transmembrane domain (aa 219-244), and a short cytoplasmic tail (aa 245-256). It enables peptide antigen binding activity and is involved in response to interferon- . It acts upstream of or within antigen processing and presentation of exogenous peptide antigen via MHC class II and plays a role in selection of TCR repertoire during positive and negative selection process and are essential for positive requirement of CD4+ cells. NIMR-4 reacts specifically with a non-polymorphic I-A-encoded epitope on MHC Class II antigens that are predominantly expressed on antigen-presenting cells, including B lymphocytes, macrophages, dendritic cells, and certain epithelial cells. (Ref.: Wosen, JE., et al. (2018). Front. Immunol. 9; 2144; Roche, PA., and Furuta, K. (2015). Nat. Rev. Immunol. 15; 203-216).

Physical form

Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.

Reconstitution

1.0 mg/mL after reconstitution at 25 μL in PBS. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C. Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial. Reconstitute each vial by adding 25 μL PBS. Protect from light.

Legal Information

ColorWheel is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.