Invitrogen

MHC Class II (I-A/I-E) Monoclonal Antibody (M5/114.15.2), Alexa Fluor™ 532, eBioscience™

Product Image Gallery

MHC Class II (I-A/I-E) Antibody in Flow Cytometry (Flow) — BALB/c mouse splenocytes were stained with CD3e Monoclonal Antibody, APC (Product # 17-0031-82) and 0.25 µg of Rat IgG2b kappa Isotype Control, Alexa Fluor 532 (Product # 58-4031-80) (left) or 0.25 µg of MHC Class II Monoclonal Antibody, Alexa Fluor 532 (right). 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

58-5321-82

Applications

Tested Dilution

Publications

Immunohistochemistry (IHC)

View 2 publications 2 publications

Flow Cytometry (Flow)

0.5 µg/test

View 19 publications 19 publications

Immunoprecipitation (IP)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Mouse

Published species

Not Applicable

Host/Isotype

Rat / IgG2b, kappa

Recommended Isotype Control

Rat IgG2b kappa Isotype Control (eB149/10H5), Alexa Fluor™ 532, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

M5/114.15.2

Conjugate

Alexa Fluor™ 532

Excitation/Emission Max

534/553 nm View spectra

Form

Liquid

Concentration

0.2 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_2811913

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 0.5 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.

Alexa Fluor 532 is excited with the Green laser (532 nm) and emits at 561 nm. This cannot be used with the Yellow-Green laser (561 nm). We recommend using a 560/14 band pass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

Excitation: 532 nm; Emission: 561 nm; Laser: Green 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.

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

Flow Cytometry

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Product References

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 # 58-5321-82) 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 # 58-5321-82) 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 # 58-5321-82) 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 # 58-5321-82) 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 # 58-5321-82) 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 # 58-5321-82) 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 # 58-5321-82) 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 # 58-5321-82) 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

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 58-5321-82) in Flow Cytometry

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 # 58-5321-82) 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

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 58-5321-82) in Flow Cytometry

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 # 58-5321-82) 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

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 58-5321-82) in Flow Cytometry

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 # 58-5321-82) 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 # 58-5321-82) 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 # 58-5321-82) 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

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 58-5321-82) in Flow Cytometry

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

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 58-5321-82) in Flow Cytometry

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

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 58-5321-82) in Flow Cytometry

Tue Aug 13 00:00:00 EDT 2019

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

Nature communications

Pascual-García M,Bonfill-Teixidor E,Planas-Rigol E,Rubio-Perez C,Iurlaro R,Arias A,Cuartas I,Sala-Hojman A,Escudero L,Martínez-Ricarte F,Huber-Ruano I,Nuciforo P,Pedrosa L,Marques C,Braña I,Garralda E,Vieito M,Squatrito M,Pineda E,Graus F,Espejo C,Sahuquillo J,Tabernero J,Seoane J

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 58-5321-82) in Flow Cytometry

Tue Jun 11 00:00:00 EDT 2019

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

Nature communications

Flores CT,Wildes TJ,Drake JA,Moore GL,Dean BD,Abraham RS,Mitchell DA

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 58-5321-82) in Flow Cytometry

Wed Oct 17 00:00:00 EDT 2018

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 # 58-5321-82) in Flow Cytometry

Thu Mar 01 00:00:00 EST 2018

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