Invitrogen

MHC Class II (I-A/I-E) Monoclonal Antibody (M5/114.15.2), Super Bright™ 780, eBioscience™

MHC Class II (I-A/I-E) Antibody in Flow Cytometry (Flow)

FIGURE: 1 / 32

MHC Class II (I-A/I-E) Antibody (78-5321-82) in Flow

Swiss Webster mouse splenocytes were stained with CD3 Monoclonal Antibody, APC (Product # 17-0031-82) and 0.125 µg of Rat IgG2b kappa Isotype Control, Super Bright 780 (Product # 78-4031-82) (left) or 0.125 µg of MHC Class II (I-A/I-E) Monoclonal Antibody, Super Bright 780 (right). Cells in the lymphocyte gate were used for analysis.

MHC Class II (I-A/I-E) Antibody in Immunohistochemistry (IHC)

MHC Class II (I-A/I-E) Antibody in Western Blot, Immunoprecipitation (WB, IP)

Product Image Gallery

Product Details

78-5321-82

Applications

Tested Dilution

Publications

Western Blot (WB)

View 1 publication 1 publication

Immunohistochemistry (IHC)

View 3 publications 3 publications

Flow Cytometry (Flow)

0.25 µg/test

View 36 publications 36 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™ 780, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

M5/114.15.2

Conjugate

Super Bright™ 780 Super Bright™ 780 Super Bright™ 780

Excitation/Emission Max

413/780 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!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2744917

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 stimulated mouse splenocytes. This may be used at less than or equal to 0.25 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 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 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.

In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).

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°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: 780 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.

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.

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Product References

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eLife

Kovacs MA,Cowan MN,Babcock IW,Sibley LA,Still K,Batista SJ,Labuzan SA,Sethi I,Harris TH

78-5321-82 was used in Flow cytometry/Cell sorting to reveal that CNS lymphatic drainage supports the development of peripheral T cell responses against T. gondii but remains dispensable for immune protection of the brain.

Wed Dec 21 00:00:00 EST 2022

Endogenous retroviruses promote homeostatic and inflammatory responses to the microbiota.

Cell

Lima-Junior DS,Krishnamurthy SR,Bouladoux N,Collins N,Han SJ,Chen EY,Constantinides MG,Link VM,Lim AI,Enamorado M,Cataisson C,Gil L,Rao I,Farley TK,Koroleva G,Attig J,Yuspa SH,Fischbach MA,Kassiotis G,Belkaid Y

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 78-5321-82) in Flow Cytometry

Thu Jul 08 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 # 78-5321-82) in Immunohistochemistry

Sat May 01 00:00:00 EDT 2021

CD11b Deficiency Exacerbates Methicillin-Resistant Staphylococcus aureus-Induced Sepsis by Upregulating Inflammatory Responses of Macrophages.

Immune network

Sim H,Jeong D,Kim HI,Pak S,Thapa B,Kwon HJ,Lee K

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 78-5321-82) in Flow Cytometry

Thu Apr 01 00:00:00 EDT 2021

Prostaglandin E2 serves a dual role in regulating the migration of dendritic cells.

International journal of molecular medicine

Diao G,Huang J,Zheng X,Sun X,Tian M,Han J,Guo J

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 78-5321-82) in Flow Cytometry

Fri Jan 01 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

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Fri Oct 29 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 # 78-5321-82) in Flow Cytometry

Mon May 24 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

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Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 78-5321-82) in Flow Cytometry

Tue Jan 12 00:00:00 EST 2021

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

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Fri Dec 10 00:00:00 EST 2021

Smek1 deficiency exacerbates experimental autoimmune encephalomyelitis by activating proinflammatory microglia and suppressing the IDO1-AhR pathway.

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Mon Jun 28 00:00:00 EDT 2021

Frequent amplification of HDAC genes and efficacy of HDAC inhibitor chidamide and PD-1 blockade combination in soft tissue sarcoma.

Journal for immunotherapy of cancer

Que Y,Zhang XL,Liu ZX,Zhao JJ,Pan QZ,Wen XZ,Xiao W,Xu BS,Hong DC,Guo TH,Shen LJ,Fan WJ,Chen HY,Weng DS,Xu HR,Zhou PH,Zhang YZ,Niu XH,Zhang X

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Mon Feb 01 00:00:00 EST 2021

Early activation of the cardiac CX3CL1/CX3CR1 axis delays β-adrenergic-induced heart failure.

Scientific reports

Flamant M,Mougenot N,Balse E,Le Fèvre L,Atassi F,Gautier EL,Le Goff W,Keck M,Nadaud S,Combadière C,Boissonnas A,Pavoine C

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Thu Sep 09 00:00:00 EDT 2021

PP2Cδ Controls the Differentiation and Function of Dendritic Cells Through Regulating the NSD2/mTORC2/ACLY Pathway.

Frontiers in immunology

Lv N,Jin S,Liang Z,Wu X,Kang Y,Su L,Dong Y,Wang B,Ma T,Shi L

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Mon Feb 28 00:00:00 EST 2022

Epinephrine Production in Th17 Cells and Experimental Autoimmune Encephalitis.

Frontiers in immunology

Yang P,Tian H,Zou YR,Chambon P,Ichinose H,Honig G,Diamond B,Kim SJ

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Mon Jul 05 00:00:00 EDT 2021

Systemic Inflammation Accelerates Changes in Microglial and Synaptic Markers in an Experimental Model of Chronic Neurodegeneration.

Frontiers in neuroscience

Chouhan JK,Püntener U,Booth SG,Teeling JL

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Sat Jan 22 00:00:00 EST 2022

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

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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

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Wed Oct 07 00:00:00 EDT 2020

E6‑regulated overproduction of prostaglandin E2 may inhibit migration of dendritic cells in human papillomavirus 16‑positive cervical lesions.

International journal of oncology

Huang J,Diao G,Zhang Q,Chen Y,Han J,Guo J

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 78-5321-82) in Flow Cytometry

Wed Apr 01 00:00:00 EDT 2020

IFN-β signalling regulates RAW 264.7 macrophage activation, cytokine production, and killing activity.

Innate immunity

Karimi Y,Giles EC,Vahedi F,Chew MV,Nham T,Loukov D,Lee AJ,Bowdish DM,Ashkar AA

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Wed Apr 01 00:00:00 EDT 2020

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

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Mon Apr 19 00:00:00 EDT 2021

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

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Mon May 04 00:00:00 EDT 2020

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

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

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Tue Apr 28 00:00:00 EDT 2020

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

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

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Fri Nov 12 00:00:00 EST 2021

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

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

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Sun Nov 29 00:00:00 EST 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

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

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Sun Apr 02 00:00:00 EDT 2023

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

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Sun Apr 02 00:00:00 EDT 2023

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International journal of molecular sciences

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

Stem cell research & therapy

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Fri Nov 06 00:00:00 EST 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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Lactobacillus reuteri 5454 and Bifidobacterium animalis ssp. lactis 5764 improve colitis while differentially impacting dendritic cells maturation and antimicrobial responses.

Scientific reports

Hrdý J,Alard J,Couturier-Maillard A,Boulard O,Boutillier D,Delacre M,Lapadatescu C,Cesaro A,Blanc P,Pot B,Ryffel B,Chamaillard M,Grangette C

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Tue Mar 24 00:00:00 EDT 2020

Nuclear imaging-guided PD-L1 blockade therapy increases effectiveness of cancer immunotherapy.

Journal for immunotherapy of cancer

Gao H,Wu Y,Shi J,Zhang X,Liu T,Hu B,Jia B,Wan Y,Liu Z,Wang F

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Sun Nov 01 00:00:00 EDT 2020

Analysis of Myeloid Cells in Mouse Tissues with Flow Cytometry.

STAR protocols

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Fri Jun 19 00:00:00 EDT 2020

CDK4/6 inhibition promotes immune infiltration in ovarian cancer and synergizes with PD-1 blockade in a B cell-dependent manner.

Theranostics

Zhang QF,Li J,Jiang K,Wang R,Ge JL,Yang H,Liu SJ,Jia LT,Wang L,Chen BL

Published figure using MHC Class II (I-A/I-E) monoclonal antibody (Product # 78-5321-82) in Flow Cytometry

Tue Jun 01 00:00:00 EDT 2021

Tregitopes regulate the tolerogenic immune response and decrease the foetal death rate in abortion-prone mouse matings.

Scientific reports

Kedzierska AE,Lorek D,Slawek A,Chelmonska-Soyta A

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Mon Jun 29 00:00:00 EDT 2020

Polyanhydride Nanoparticles Induce Low Inflammatory Dendritic Cell Activation Resulting in CD8⁺ T Cell Memory and Delayed Tumor Progression.

International journal of nanomedicine

Darling R,Senapati S,Christiansen J,Liu L,Ramer-Tait AE,Narasimhan B,Wannemuehler M

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Mon Dec 07 00:00:00 EST 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

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

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Tue Jun 11 00:00:00 EDT 2019

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

Experimental and therapeutic medicine

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

Nature communications

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Wed Oct 17 00:00:00 EDT 2018

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

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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

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UniProt ID: (Mouse) P14434, (Mouse) P14483

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Entrez Gene ID: (Mouse) 14960, (Mouse) 14961, (Mouse) 14969

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Function(s)

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

Process(es)

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

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MHC Class II (I-A/I-E) Monoclonal Antibody (M5/114.15.2), Super Bright™ 780, eBioscience™

MHC Class II (I-A/I-E) Antibody (78-5321-82) in Flow

Product Details

78-5321-82

Western Blot (WB)

Immunohistochemistry (IHC)

Flow Cytometry (Flow)

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

Shipping conditions

RRID

Product Specific Information

Target Information

How to use the Panel Builder

Bioinformatics

Performance Guarantee

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