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Invitrogen
MHC Class II (I-A/I-E) Monoclonal Antibody (M5/114.15.2), NovaFluor™ Yellow 660, eBioscience™
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FIGURE: 1 / 4
MHC Class II (I-A/I-E) Antibody (M024T02Y04-A) in Flow
C57BL/6 mouse splenocytes were unstained (left) or stained with 0.4 µg of MHC Class II (I-A/I-E) Monoclonal Antibody, NovaFluor Yellow 660 (right). All cells were co-stained with CD3e (145-2C18) Monoclonal Antibody, eFluor 450 (Product # 48-0031-89). Total viable cells were used for analysis, as determined by LIVE/DEAD Blue (Product # L34969). Data was acquired on a 5-laser Cytek Aurora and unmixed with aut... View More
Product Details
M024T02Y04-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG2b, kappa
Class
Monoclonal
Type
Antibody
Clone
M5/114.15.2
Conjugate
Excitation/Emission Max
550/664 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
AB_3099028 (AB_2910837)
Product Specific Information
Description
The M5/114.15.2 monoclonal antibody reacts with the mouse major histocompatibility complex class II, both I-A and I-E subregion-encoded glycoproteins (I-A b, I-A d, I-A q, I-E d, I-E k, not I-A f, I-A k, or I-A s). It detects a polymorphic determinant present on B cells, monocytes, macrophages, dendritic cells, and activated T lymphocytes from mice carrying the H-2 b, H-2 d, H-2 q, H-2 p, H-2 r and H-2 u but not from mice carrying the H-2 s or H-2 f haplotypes. The M5/114 mAb is reported to inhibit I-A-restricted T cell responses of the H-2 b, H-2 d, H-2 q, H-2 u but not H-2 f, H-2 k, or H-2 s haplotypes.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This M5/114.15.2 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.4 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Our internal testing shows that NovaFluor Yellow 660 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
Excitation: 550 nm; Emission: 664 nm; Laser: 561 nm (Yellow) Laser
Target Information
MHC (major histocompatibility complex) class II molecules are transmembrane glycoproteins expressed on the surface of professional antigen-presenting cells, such as macrophages, dendritic cells and B cells. Before their exposition on the cell surface, the MHC class II molecules react with endocytosed exogenous antigens, which are then presented to the T cells. The antigen-binding grove between MHC class II alpha and beta chain is open at both ends and is 15-24 amino acid residues long.
HLA and MHC antibodies play a significant role in Immunopeptidomics, facilitating the identification and characterization of neoantigens through high-performance liquid chromatography coupled to tandem Mass Spectrometry.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
How to use the Panel Builder
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
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Bioinformatics
Protein Aliases: A alpha; cell surface glycoprotein; H-2 class II histocompatibility antigen, A beta chain; H-2 class II histocompatibility antigen, A-B alpha chain; H-2 class II histocompatibility antigen, A-F alpha chain; H-2 class II histocompatibility antigen, A-K alpha chain; H-2 class II histocompatibility antigen, A-Q alpha chain; H-2 class II histocompatibility antigen, A-R alpha chain; H-2 class II histocompatibility antigen, A-S alpha chain; H-2 class II histocompatibility antigen, A-U alpha chain; H-2 class II histocompatibility antigen, I-E beta chain; histocompatibility 2, class II antigen A, alpha; histocompatibility 2, class II antigen A, beta 1; I-Ab; I-Ad; I-Aq; I-E beta MHC class II; I-Ed; I-Ek; IAalpha; major histocompatibility complex class II beta chain; major histocompatibility complex H-2E beta chain; major histocompatibility protein class II alpha chain; MHC class 2; MHC class II antigen A alpha isoform-004; MHC class II antigen A beta; MHC class II H2-IA-beta-psi; MHC class II IE-b; MHC H2-IE-beta cell surface glycoprotein; MHC-II I-A(d) alpha; response to metastatic cancers 1
Gene Aliases: Aalpha; Abeta; AI845868; H-2Aa; H-2Ab; H-2Eb; H2-Aa; H2-Ab; H2-Ab1; H2-iabeta; H2Aa; H2Eb; I-Aalpha; I-Abeta; Ia-1; Ia-2; Ia-4; Ia1; Ia2; Ia4; IAalpha; IAb; Rmcs1
UniProt ID: (Mouse) P14434, (Mouse) P14483
Entrez Gene ID: (Mouse) 14960, (Mouse) 14961, (Mouse) 14969
immune system process
antigen processing and presentation of peptide or polysaccharide antigen via MHC class II
immune response
antigen processing and presentation
antigen processing and presentation of exogenous peptide antigen via MHC class II
response to interferon-gamma
negative regulation of T cell proliferation
positive regulation of T cell differentiation
antigen processing and presentation of peptide antigen
B cell affinity maturation
immunoglobulin production involved in immunoglobulin mediated immune response
humoral immune response mediated by circulating immunoglobulin
positive regulation of antigen processing and presentation
positive regulation of T-helper 1 type immune response
positive regulation of alpha-beta T cell activation
cellular response to interferon-gamma
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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