Anti mhc class 2 m5 114

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Anti-MHC Class II (M5/114.15.2) is a laboratory reagent that binds to major histocompatibility complex (MHC) class II molecules. It is commonly used as a research tool in immunology studies.

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MHC class II (12 citations) Anti-MHC class II (5 citations) M5/114.15.2 (4 citations) MHC-II (4 citations) Anti-MHC-II (2 citations)

3 protocols using anti mhc class 2 m5 114

Immunostaining Protocols for PLET1 and Cytokeratins

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MTS24 (IgG2a) [27] (link), a rat mAb that recognises PLET1 [28] (link) was a kind gift from R.L. Boyd; 1D4 (anti-PLET1, rat IgG, [28] (link)), anti-Cytokeratin 8 (Troma 1, rat IgG2a, DSHB); anti-Cytokeratin 14 (AF64, rabbit IgG, Covance)); anti-Cytokeratin 5 (AF138, rabbit IgG, Covance); anti-CD4-FITC or PE (H129.19, rat IgG2a); anti-CD8-FITC (53–6.7, rat IgG2a); anti-CD11b-FITC (M1/70, rat IgG2b); anti-CD19-FITC (1D3, rat IgG2a); anti-CD25-PE (3C7, rat IgG2b); anti-CD44-APC (1M7, rat IgG2b); anti-Ly76-FITC (Ter119, rat IgG2b); anti-CD45-APC (30-F11, rat IgG2b); anti-Cytokeratin (rabbit IgG polyclonal, DAKO); biotinlyated UEA-1 (Lectin, Vector Laboratories); anti-MHC Class II (M5/114.15.2, rat IgG2b, BD Bioscience); anti-AIRE (M-300, SCBT); anti-CD205 (NLDC-145, AbD Serotec); (CDR1 (CDR1, rat IgG2a, Gift from B Kyewski). For detection of unconjugated primaries the following secondary antibodies were used; goat anti-rabbit IgG-alexa488; goat anti-rat IgG-alexa647; Streptavidin-alexa647, goat anti-rat IgG-alexa568 (all Molecular Probes).

Jin X., Nowell C.S., Ulyanchenko S., Stenhouse F.H, & Blackburn C.C. (2014). Long-Term Persistence of Functional Thymic Epithelial Progenitor Cells In Vivo under Conditions of Low FOXN1 Expression. PLoS ONE, 9(12), e114842.

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Multiparametric Flow Cytometry Analysis

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The harvested spleens were mashed and single-cell suspensions of the spleens were stained with the following surface antibodies: anti-CD45 (30-F11, BD Biosciences), LIVE/DEAD Fixable Blue cell stain (Invitrogen), anti-CD19 (ID3, BD Biosciences), anti-CD8a (53-6.7, BioLegend), anti-CD27 (LG7F9, eBioscience), anti-CD11b (M1/70, BioLegend), anti-CD11c (HL3, BD Biosciences), anti-CD86 (GL1, BioLegend), anti-NK1.1 (PK136, BioLegend), anti-NKp46 (29A1.4, BioLegend), anti-MHC Class II (M5/114.15.2, BD Biosciences), anti-F4/80 (BM8, BioLegend), anti-CD80 (16-10A1), anti-CD3 (17A2, BioLegend), anti-CD4 (RM4-5, Invitrogen), anti-CTLA-4 (UC10-4B9, BioLegend), anti-PD-1 (29F.1A12, eBioscience), anti-CD28 (37.51, BioLegend), anti-CD44 (IM7, BD Biosciences), anti-CD43 (1B11, BioLegend), anti-CD47 (miap301, BioLegend), anti-CD62L (MEL-14, BioLegend), anti-CD25 (PC61.5, eBioscience), and anti-CD107a (1D4B, BioLegend). Intracellular staining was then performed with a FOXP3 permeabilization and fixation kit (eBioscience) according to manufacturer’s recommendations using the following antibodies: Ki-67 (B56; BD Biosciences) and GrB (QA16A02, BioLegend). Flow cytometric data were collected with a Beckman Coulter Cytoflex LX (6-L NUV) flow cytometer and analyzed using FlowJo software (Tree Star).

Rao D., O’Donnell K.L., Carmody A., Weissman I.L., Hasenkrug K.J, & Marzi A. (2021). CD47 expression attenuates Ebola virus-induced immunopathology in mice. Antiviral research, 197, 105226.

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Skin Sensitization and Migratory DC Analysis

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Mice were sensitized with 150 µL of 1.0% FITC (DOJINDO, Kumamoto, Japan) in acetone/dibutyl phthalate (1:1, v/v) was applied on shaved dorsal skin at day 0.
Migratory DCs were analyzed as previously described^{34 (link)}. Flow cytometric experiments were conducted on draining LN cells collected from the axillary/inguinal LNs one day after sensitization. For single-cell suspensions, preparation from the draining LNs was digested with collagenase type I (Sigma-Aldrich, 400 U/L) at 37 °C for 30 min and incubated with ethylenediaminetetraacetic acid (EDTA, final concentration 5 mM) for an additional 10 min. Cell suspensions were Fc blocked with CD16/CD32 (2.4G2, BD Bioscience) and stained with the following anti-mouse antibodies purchased from BD Bioscience: anti-MHC class II (M5/114.15.2) and anti-CD86 (GL1). Dead cells were labeled with 7-AAD (BD Biosciences).

Tetsu H., Nakayama K., Nishijo T., Yuki T, & Miyazawa M. (2023). CTLA-4 suppresses hapten-induced contact hypersensitivity in atopic dermatitis model mice. Scientific Reports, 13, 7936.

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