Invitrogen

MHC Class II (I-A/I-E) Monoclonal Antibody (M5/114.15.2), Super Bright™ 600, eBioscience™

Product Image Gallery

MHC Class II (I-A/I-E) Antibody in Flow Cytometry (Flow) — C57BL/6 mouse splenocytes were stained with CD3 Monoclonal Antibody, FITC (Product # 11-0031) and 0.06 µg of MHC Class II (I-A/I-E) Monoclonal Antibody, Super Bright 600. Cells in the lymphocyte gate were used for analysis.

MHC Class II (I-A/I-E) Antibody in Immunohistochemistry (IHC) — Figure 5 The entry to the brain of rAAV9 with MRIgFUS and PHP.B coincides with markers of an adaptive immune response Brain sections were immunostained for MHC class II, the T cell marker CD3, and GFP following treatment with MRIgFUS and injection of 1E11AAV9 (A), 1E10PHP.B (B), and 1E11PHP.B (C). Insets in (A), (B), and (C) correspond to (A1-A5), (B1-B4), and (C1-C4), respectively. Insets in (A1-A5), (B1-B4), and (C1-C4) correspond to (a1-a5), (b1-b4), and (c1-c4), respectively. T cell infiltration and MHC class II expression are visible in MRIgFUS spots in animals injected with rAAV9 but not in the contralateral site and in multiple brain areas in animals injected with 1E11PHP.B but not 1E10PHP.B. MHC class II expression is primarily seen around large vessels, and T cells also infiltrate into the brain parenchyma. The number of T cells per square millimeter is displayed for each mouse and each MRIgFUS spot (for rAAV9; D) or brain area (for PHP.B; E). Quantifications show variability in T cell invasion within and between mice in both groups, as well as T cell invasion in all brain areas quantified in mice injected with 1E11PHP.B. Ctx, cortex; Str, striatum; Th, thalamus; HC, hippocampus. 5,000 mum scale bar (C) applies to (A-C), 250 mum scale bar (C4) applies to (A1-C4), and 50 mum scale bar (c4) applies to (a1-c4).

MHC Class II (I-A/I-E) Antibody in Immunoprecipitation (IP) — Figure 3 Inhibition of RON leads to increased transcription of March-I without affecting March-8, resulting in increased MHC II and CD86 ubiquitination. (A) DCs were stimulated for 4 h and total mRNA was collected using an RNeasy mini column kit. The level of March-I and March-8 mRNA was tested by RT-PCR and GUSB expression was used as an internal control. The data represent the mean +- SEM of three independent experiments. NS, No significant difference; *p < 0.05; **p < 0.01. Heat map of gene expression for March-1 and March-8, comparing unstimulated DCs are shown. (B) DCs were stimulated for 4 h and 1 uM MG132 was added at the same time. The immune precipitates were analyzed by immunoblotting using anti-ubiquitin antibodies and the secondary antibody was anti-mouse IgG for immunoprecipitation to eliminate the effects of light and heavy chains. The experiment was repeated twice independently.

Product Details

63-5321-82

Applications	Tested Dilution	Publications
Flow Cytometry (Flow)	0.125 µg/test	View 20 publications 20 publications
Immunohistochemistry (IHC)	-	View 2 publications 2 publications
Immunoprecipitation (IP)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Mouse
Published species	Mouse
Host/Isotype	Rat / IgG2b, kappa
Recommended Isotype Control	Rat IgG2b kappa Isotype Control (eB149/10H5), Super Bright™ 600, eBioscience™
Class	Monoclonal
Type	Antibody
Clone	M5/114.15.2
Conjugate	Super Bright™ 600 Alexa Fluor 532 Alexa Fluor 700 APC APC-eFluor 780 Biotin Brilliant UV 496 Brilliant UV 661 Brilliant UV 737 Brilliant Violet 421 Brilliant Violet 650 Brilliant Violet 711 Brilliant Violet 786 eFluor 450 eFluor 506 FITC Functional Grade NovaFluor Blue 510 NovaFluor Blue 610-70S NovaFluor Blue 660-120S NovaFluor Red 685 NovaFluor Red 710 NovaFluor Yellow 570 NovaFluor Yellow 590 NovaFluor Yellow 660 PE PE-Cyanine5 PE-Cyanine7 PE-eFluor 610 PerCP-eFluor 710 Super Bright 436 Super Bright 645 Super Bright 702 Super Bright 780 Unconjugated
Excitation/Emission Max	414/601 nm View spectra
Form	Liquid
Concentration	0.2 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2, with BSA
Contains	0.09% sodium azide
Storage conditions	4° C, store in dark, DO NOT FREEZE!
RRID	AB_2688076

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.

Applications Reported: This M5/114.15.2 antibody has been reported for use in flow cytometric analysis.

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 0.125 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.

Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4 degrees C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.

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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Flow Cytometry

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Product References

Neoantigen vaccine-induced CD4 T cells confer protective immunity in a mouse model of multiple myeloma through activation of CD8 T cells against non-vaccine, tumor-associated antigens.

Journal for immunotherapy of cancer

Bekri S,Rodney-Sandy R,Gruenstein D,Mei A,Bogen B,Castle J,Levey D,Cho HJ

63-5321-82 was used in Flow cytometry/Cell sorting to indicate that activation of CD8 CTL against MOPC315-associated antigens not present in the vaccine is one of the major mechanisms of tumor immunity.

Tue Feb 01 00:00:00 EST 2022

Transgene distribution and immune response after ultrasound delivery of rAAV9 and PHP.B to the brain in a mouse model of amyloidosis.

Molecular therapy. Methods & clinical development

Kofoed RH,Heinen S,Silburt J,Dubey S,Dibia CL,Maes M,Simpson EM,Hynynen K,Aubert I

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Immunocytochemistry

Fri Dec 10 00:00:00 EST 2021

MiR-103 protects from recurrent spontaneous abortion via inhibiting STAT1 mediated M1 macrophage polarization.

International journal of biological sciences

Zhu X,Liu H,Zhang Z,Wei R,Zhou X,Wang Z,Zhao L,Guo Q,Zhang Y,Chu C,Wang L,Li X

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Fri Nov 12 00:00:00 EST 2021

Wip1 regulates the immunomodulatory effects of murine mesenchymal stem cells in type 1 diabetes mellitus via targeting IFN-α/BST2.

Cell death discovery

Zhou N,Liu W,Zhang W,Liu Y,Li X,Wang Y,Zheng R,Zhang Y

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Fri Oct 29 00:00:00 EDT 2021

RON Expression Mediates Lipopolysaccharide-Mediated Dendritic Cell Maturation via March-I.

Frontiers in cellular and infection microbiology

Huang L,Fang X,Zhang X,Wu W,Yao H,Fang Q

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Immunoprecipitation

Thu Jun 17 00:00:00 EDT 2021

Topical arginase inhibition decreases growth of cutaneous squamous cell carcinoma.

Scientific reports

Mittal A,Wang M,Vidyarthi A,Yanez D,Pizzurro G,Thakral D,Tracy E,Colegio OR

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Mon May 24 00:00:00 EDT 2021

Regionally diverse astrocyte subtypes and their heterogeneous response to EAE.

Glia

Borggrewe M,Grit C,Vainchtein ID,Brouwer N,Wesseling EM,Laman JD,Eggen BJL,Kooistra SM,Boddeke EWGM

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Immunohistochemistry

Sat May 01 00:00:00 EDT 2021

Dietary Glucose Consumption Promotes RALDH Activity in Small Intestinal CD103⁺CD11b⁺ Dendritic Cells.

Frontiers in immunology

Ko HJ,Hong SW,Verma R,Jung J,Lee M,Kim N,Kim D,Surh CD,Kim KS,Rudra D,Im SH

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Mon Apr 19 00:00:00 EDT 2021

Targeting Phosphatidylserine Enhances the Anti-tumor Response to Tumor-Directed Radiation Therapy in a Preclinical Model of Melanoma.

Cell reports

Budhu S,Giese R,Gupta A,Fitzgerald K,Zappasodi R,Schad S,Hirschhorn D,Campesato LF,De Henau O,Gigoux M,Liu C,Mazo G,Deng L,Barker CA,Wolchok JD,Merghoub T

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Tue Jan 12 00:00:00 EST 2021

Smooth muscle-derived progenitor cell myofibroblast differentiation through KLF4 downregulation promotes arterial remodeling and fibrosis.

JCI insight

Lu S,Jolly AJ,Strand KA,Dubner AM,Mutryn MF,Moulton KS,Nemenoff RA,Majesky MW,Weiser-Evans MC

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Thu Dec 03 00:00:00 EST 2020

Immunomodulatory Effects of Recombinant Mycobacterium smegmatis Expressing Antigen-85B Epitopes in Infected J774A.1 Murine Macrophages.

Pathogens (Basel, Switzerland)

Kadir NA,Acosta A,Sarmiento ME,Norazmi MN

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Sun Nov 29 00:00:00 EST 2020

Skeletal muscle healing by M1-like macrophages produced by transient expression of exogenous GM-CSF.

Stem cell research & therapy

Martins L,Gallo CC,Honda TSB,Alves PT,Stilhano RS,Rosa DS,Koh TJ,Han SW

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Fri Nov 06 00:00:00 EST 2020

Glufosinate constrains synchronous and metachronous metastasis by promoting anti-tumor macrophages.

EMBO molecular medicine

Menga A,Serra M,Todisco S,Riera-Domingo C,Ammarah U,Ehling M,Palmieri EM,Di Noia MA,Gissi R,Favia M,Pierri CL,Porporato PE,Castegna A,Mazzone M

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Wed Oct 07 00:00:00 EDT 2020

Blockade of the AHR restricts a Treg-macrophage suppressive axis induced by L-Kynurenine.

Nature communications

Campesato LF,Budhu S,Tchaicha J,Weng CH,Gigoux M,Cohen IJ,Redmond D,Mangarin L,Pourpe S,Liu C,Zappasodi R,Zamarin D,Cavanaugh J,Castro AC,Manfredi MG,McGovern K,Merghoub T,Wolchok JD

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Tue Aug 11 00:00:00 EDT 2020

Optimized Protocol for Isolation of Small Extracellular Vesicles from Human and Murine Lymphoid Tissues.

International journal of molecular sciences

Bordas M,Genard G,Ohl S,Nessling M,Richter K,Roider T,Dietrich S,Maaß KK,Seiffert M

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Tue Aug 04 00:00:00 EDT 2020

Diversity in medullary thymic epithelial cells controls the activity and availability of iNKT cells.

Nature communications

Lucas B,White AJ,Cosway EJ,Parnell SM,James KD,Jones ND,Ohigashi I,Takahama Y,Jenkinson WE,Anderson G

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Mon May 04 00:00:00 EDT 2020

Ly6c^Lo non-classical monocytes promote resolution of rhesus rotavirus-mediated perinatal hepatic inflammation.

Scientific reports

Alkhani A,Levy CS,Tsui M,Rosenberg KA,Polovina K,Mattis AN,Mack M,Van Dyken S,Wang BM,Maher JJ,Nijagal A

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

Tue Apr 28 00:00:00 EDT 2020

Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms.

Journal for immunotherapy of cancer

Neuwelt AJ,Kimball AK,Johnson AM,Arnold BW,Bullock BL,Kaspar RE,Kleczko EK,Kwak JW,Wu MH,Heasley LE,Doebele RC,Li HY,Nemenoff RA,Clambey ET

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Wed Apr 01 00:00:00 EDT 2020

Artificial Mini Dendritic Cells Boost T Cell-Based Immunotherapy for Ovarian Cancer.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Cheng S,Xu C,Jin Y,Li Y,Zhong C,Ma J,Yang J,Zhang N,Li Y,Wang C,Yang Z,Wang Y

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Wed Apr 01 00:00:00 EDT 2020

cGAS/STING axis mediates a topoisomerase II inhibitor-induced tumor immunogenicity.

The Journal of clinical investigation

Wang Z,Chen J,Hu J,Zhang H,Xu F,He W,Wang X,Li M,Lu W,Zeng G,Zhou P,Huang P,Chen S,Li W,Xia LP,Xia X

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LIF regulates CXCL9 in tumor-associated macrophages and prevents CD8⁺ T cell tumor-infiltration impairing anti-PD1 therapy.

Nature communications

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Lin^-CCR2⁺ hematopoietic stem and progenitor cells overcome resistance to PD-1 blockade.

Nature communications

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Effect of immune tolerance induced by immature dendritic cells and CTLA4-Ig on systemic lupus erythematosus: An in vivo study.

Experimental and therapeutic medicine

Huang C,Zhang L,Ling F,Wen S,Luo Y,Liu H,Liu J,Zheng W,Liang M,Sun J,Lin YK

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 63-5321-82) in Flow Cytometry

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Bioinformatics

Protein Aliases: cell surface glycoprotein; H-2 class II histocompatibility antigen, A beta chain; H-2 class II histocompatibility antigen, I-E beta chain; histocompatibility 2, class II antigen A, beta 1; I-Ab; I-Ad; I-Aq; I-E beta MHC class II; I-Ed; I-Ek; major histocompatibility complex class II beta chain; major histocompatibility complex H-2E beta chain; MHC class 2; 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; response to metastatic cancers 1

Gene Aliases: Abeta; AI845868; H-2Ab; H-2Eb; H2-Ab; H2-Ab1; H2-iabeta; H2Eb; I-Abeta; Ia-2; Ia-4; Ia2; Ia4; IAb; Rmcs1

UniProt ID: (Mouse) P14483

Entrez Gene ID: (Mouse) 14961, (Mouse) 14969

Molecular Function: major histocompatibility complex protein

Function(s)

B cell affinity maturation immune system process immunoglobulin production involved in immunoglobulin mediated immune response humoral immune response mediated by circulating immunoglobulin antigen processing and presentation of peptide or polysaccharide antigen via MHC class II positive regulation of antigen processing and presentation positive regulation of T-helper 1 type immune response immune response antigen processing and presentation antigen processing and presentation of exogenous peptide antigen via MHC class II negative regulation of T cell proliferation positive regulation of alpha-beta T cell activation antigen processing and presentation of peptide antigen cellular response to interferon-gamma response to interferon-gamma

Process(es)

protein binding transcription factor binding toxic substance binding ubiquitin protein ligase binding peptide antigen binding protein heterodimerization activity repressing transcription factor binding

MHC Class II (I-A/I-E) Monoclonal Antibody (M5/114.15.2), Super Bright™ 600, eBioscience™

Product Details

63-5321-82

Flow Cytometry (Flow)

Immunohistochemistry (IHC)

Immunoprecipitation (IP)

Species Reactivity

Published species

Host/Isotype

Recommended Isotype Control

Class

Type

Clone

Conjugate

Excitation/Emission Max

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

How to use the Panel Builder

Bioinformatics

Performance Guarantee

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