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cell lines b16f10 β2m  (ATCC)


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    ATCC cell lines b16f10 β2m
    Cell Lines B16f10 β2m, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 7965 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/%CE%B22m/B16-F10/pm41758652-249-82-112
    Average 99 stars, based on 7965 article reviews
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    Western Blot:

    Article Title: Distinct differences in association of MHC class I with endoplasmic reticulum proteins in wild-type, and beta 2-microglobulin- and TAP-deficient cell lines.
    Article Snippet: The cell lines were cultured in RPMI 1640 medium (Gibco/BRL, Gaithersburg, MD), supple- For sequential immunoprecipitation, the first round immunoprecipitates were disrupted by incubating in 0.5 ml of 0.2%mented with 10% heat-inactivated FBS, 100 U/ml penicillin, 100 μg/ml streptomycin and 2 mM glutamine, at 37°C in a SDS at 37°C for 1 h. The samples were then diluted to 5 ml in 1% NP-40 lysis buffer and the second antiserum was added5% CO2 atmosphere. for a second round of immunoprecipitation. .. Antibodies For Western immunoblotting, aliquots of cell lysates, or immunoprecipitates, were loaded onto a 10% SDS–PAGE.mAb W6/32, specific to β2m-associated human class I molecules (43), was obtained from the ATCC (Rockville, Proteins were transferred onto a nitrocellulose filter, which was probed with the anti-BiP antibody at a dilution of 1:500,MD). ..

    Membrane:

    Article Title: Degradation of Transcription Factor Rfx5 during the Inhibition of Both Constitutive and Interferon γ–Inducible Major Histocompatibility Complex Class I Expression in Chlamydia-Infected Cells
    Article Snippet: Rabbit antibodies were used to detect RFX5 (Rockland Immunochemicals), USF-1 (SC-229; Santa Cruz Biotechnology, Inc.), USF-2 (SC-862; Santa Cruz), and 11S regulatory subunit PA28α (PW8185; AFFINITI Research Products Ltd.). .. Mouse antibodies were used to detect HLA-A and -B (HB296; ATCC), β2M (HB149; ATCC), β1 integrin (provided by Dr. J. Wilkins, University of Manitoba, Manitoba, Canada), 20S proteasome α subunit HC2 (MCP20; AFFINITI), 20S proteasome α subunit HC3 (MCP21; AFFINITI), 20S proteasome subunits α1, 2, 3, 5, 6, and 7 (MCP231; AFFINITI), 20S proteasome subunit β7 (MCP205; AFFINITI), and a chlamydial major outer membrane protein (MOMP; clone MC22; our unpublished data). .. Primary antibody binding was detected with horseradish peroxidase–conjugated goat anti–mouse IgG or –rabbit IgG, depending on the source of the primary antibodies, and visualized using an ECL kit (Amersham).



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    cell lines b16f10 β2m  (ATCC)
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    Cell Lines B16f10 β2m, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human beta 2 microglobulin β2m elisa pair set  (Sino Biological)
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    a , Top: schematic of double-positive thymocytes from CD8 Dual mice. The altered Cd4 gene locus encodes CD8α.1 co-receptors (composed of CD8α.1CD8β dimers), and the endogenous Cd8 gene locus encodes CD8α.2 co-receptors (composed of CD8α.2CD8β dimers). Bottom: schematic of the altered Cd4 CD8 and endogenous Cd8 CD8 genes in CD8 Dual mice; IRES, internal ribosome entry site; poly A, polyadenylation signals. b , Flow cytometry analysis of whole thymocytes from CD8 Dual ( n = 16) and B6 ( n = 15) mice (representative of 15 independent experiments). Top: Cd4 -encoded co-receptors (CD8α.1 or CD4) versus Cd8 -encoded co-receptor CD8α.2 profile, showing four thymocyte subsets; double-negative (DN), double-positive (DP) and two single-positive T cell subsets. Total cell number (mean ± s.e.m) is shown above profiles. Middle: TCRβ histogram, showing TCRβ hi (TCR hi ) thymocytes. Bottom: CD24 versus TCRβ profile, showing CD24 − TCR + mature thymocytes. c , Flow cytometry analysis of TCR hi thymocytes and TCR + LN T cells from CD8 Dual ( n = 16) and B6 ( n = 15) mice, showing Cd4 -encoded co-receptor (CD8α.1 or CD4) versus Cd8 -encoded co-receptor (CD8α.2) or CD8β profiles. TCR expression of LN cells are shown in Extended Data Fig. . Numbers (mean ± s.e.m.) of TCR hi thymocytes and LN T cells are shown above profiles (representative of 15 independent experiments). d , Numbers of CD8β + T cells among CD24 − TCR + mature thymocytes (T) and TCR + LN T cells (L) in CD8 Dual mice with the indicated MHC deficiencies. CD8β histograms are shown in Extended Data Fig. (WT: T ( n = 6), L ( n = 9), MHC-II KO : T ( n = 9), L ( n = 9), <t>β2m</t> KO : T ( n = 7), L ( n = 9), MHC KO : T ( n = 3), L ( n = 5), 3–7 independent experiments). e , Mean fluorescence intensity (MFI) of Runx3d-YFP and ThPOK-GFP reporter expression in CD8.1, CD8.2, CD4 + and CD8 + T cells among TCR hi thymocytes and TCR + LN T cells in CD8 Dual and B6 mice based on histograms shown in Extended Data Fig. (Runx3d-YFP: CD8 Dual T ( n = 4), L ( n = 4), B6 T ( n = 5), L ( n = 4), ThPOK-GFP: CD8 Dual T ( n = 3), L ( n = 3), B6 T ( n = 4) L ( n = 3), representative of 3–5 independent experiments). f , RNA-seq analysis of CD62L + TCR + LN T cells from CD8 Dual and B6 mice. Genes differentially expressed between B6 CD4 + and CD8 + LN T cells were evaluated in the heat map for analysis of gene expression in CD8.1 and CD8.2 LN T cells from CD8 Dual mice ( n = 3 per group, P < 0.05, 5-fold change). Numbers within profiles and histograms indicate frequency of cells in each box or lines ( b and c ). *** P < 0.001, ** P < 0.01, * P < 0.05 (two-tailed unpaired t -tests); mean ± s.e.m. ( d and e ).
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    a , Top: schematic of double-positive thymocytes from CD8 Dual mice. The altered Cd4 gene locus encodes CD8α.1 co-receptors (composed of CD8α.1CD8β dimers), and the endogenous Cd8 gene locus encodes CD8α.2 co-receptors (composed of CD8α.2CD8β dimers). Bottom: schematic of the altered Cd4 CD8 and endogenous Cd8 CD8 genes in CD8 Dual mice; IRES, internal ribosome entry site; poly A, polyadenylation signals. b , Flow cytometry analysis of whole thymocytes from CD8 Dual ( n = 16) and B6 ( n = 15) mice (representative of 15 independent experiments). Top: Cd4 -encoded co-receptors (CD8α.1 or CD4) versus Cd8 -encoded co-receptor CD8α.2 profile, showing four thymocyte subsets; double-negative (DN), double-positive (DP) and two single-positive T cell subsets. Total cell number (mean ± s.e.m) is shown above profiles. Middle: TCRβ histogram, showing TCRβ hi (TCR hi ) thymocytes. Bottom: CD24 versus TCRβ profile, showing CD24 − TCR + mature thymocytes. c , Flow cytometry analysis of TCR hi thymocytes and TCR + LN T cells from CD8 Dual ( n = 16) and B6 ( n = 15) mice, showing Cd4 -encoded co-receptor (CD8α.1 or CD4) versus Cd8 -encoded co-receptor (CD8α.2) or CD8β profiles. TCR expression of LN cells are shown in Extended Data Fig. . Numbers (mean ± s.e.m.) of TCR hi thymocytes and LN T cells are shown above profiles (representative of 15 independent experiments). d , Numbers of CD8β + T cells among CD24 − TCR + mature thymocytes (T) and TCR + LN T cells (L) in CD8 Dual mice with the indicated MHC deficiencies. CD8β histograms are shown in Extended Data Fig. (WT: T ( n = 6), L ( n = 9), MHC-II KO : T ( n = 9), L ( n = 9), <t>β2m</t> KO : T ( n = 7), L ( n = 9), MHC KO : T ( n = 3), L ( n = 5), 3–7 independent experiments). e , Mean fluorescence intensity (MFI) of Runx3d-YFP and ThPOK-GFP reporter expression in CD8.1, CD8.2, CD4 + and CD8 + T cells among TCR hi thymocytes and TCR + LN T cells in CD8 Dual and B6 mice based on histograms shown in Extended Data Fig. (Runx3d-YFP: CD8 Dual T ( n = 4), L ( n = 4), B6 T ( n = 5), L ( n = 4), ThPOK-GFP: CD8 Dual T ( n = 3), L ( n = 3), B6 T ( n = 4) L ( n = 3), representative of 3–5 independent experiments). f , RNA-seq analysis of CD62L + TCR + LN T cells from CD8 Dual and B6 mice. Genes differentially expressed between B6 CD4 + and CD8 + LN T cells were evaluated in the heat map for analysis of gene expression in CD8.1 and CD8.2 LN T cells from CD8 Dual mice ( n = 3 per group, P < 0.05, 5-fold change). Numbers within profiles and histograms indicate frequency of cells in each box or lines ( b and c ). *** P < 0.001, ** P < 0.01, * P < 0.05 (two-tailed unpaired t -tests); mean ± s.e.m. ( d and e ).
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    a , Top: schematic of double-positive thymocytes from CD8 Dual mice. The altered Cd4 gene locus encodes CD8α.1 co-receptors (composed of CD8α.1CD8β dimers), and the endogenous Cd8 gene locus encodes CD8α.2 co-receptors (composed of CD8α.2CD8β dimers). Bottom: schematic of the altered Cd4 CD8 and endogenous Cd8 CD8 genes in CD8 Dual mice; IRES, internal ribosome entry site; poly A, polyadenylation signals. b , Flow cytometry analysis of whole thymocytes from CD8 Dual ( n = 16) and B6 ( n = 15) mice (representative of 15 independent experiments). Top: Cd4 -encoded co-receptors (CD8α.1 or CD4) versus Cd8 -encoded co-receptor CD8α.2 profile, showing four thymocyte subsets; double-negative (DN), double-positive (DP) and two single-positive T cell subsets. Total cell number (mean ± s.e.m) is shown above profiles. Middle: TCRβ histogram, showing TCRβ hi (TCR hi ) thymocytes. Bottom: CD24 versus TCRβ profile, showing CD24 − TCR + mature thymocytes. c , Flow cytometry analysis of TCR hi thymocytes and TCR + LN T cells from CD8 Dual ( n = 16) and B6 ( n = 15) mice, showing Cd4 -encoded co-receptor (CD8α.1 or CD4) versus Cd8 -encoded co-receptor (CD8α.2) or CD8β profiles. TCR expression of LN cells are shown in Extended Data Fig. . Numbers (mean ± s.e.m.) of TCR hi thymocytes and LN T cells are shown above profiles (representative of 15 independent experiments). d , Numbers of CD8β + T cells among CD24 − TCR + mature thymocytes (T) and TCR + LN T cells (L) in CD8 Dual mice with the indicated MHC deficiencies. CD8β histograms are shown in Extended Data Fig. (WT: T ( n = 6), L ( n = 9), MHC-II KO : T ( n = 9), L ( n = 9), <t>β2m</t> KO : T ( n = 7), L ( n = 9), MHC KO : T ( n = 3), L ( n = 5), 3–7 independent experiments). e , Mean fluorescence intensity (MFI) of Runx3d-YFP and ThPOK-GFP reporter expression in CD8.1, CD8.2, CD4 + and CD8 + T cells among TCR hi thymocytes and TCR + LN T cells in CD8 Dual and B6 mice based on histograms shown in Extended Data Fig. (Runx3d-YFP: CD8 Dual T ( n = 4), L ( n = 4), B6 T ( n = 5), L ( n = 4), ThPOK-GFP: CD8 Dual T ( n = 3), L ( n = 3), B6 T ( n = 4) L ( n = 3), representative of 3–5 independent experiments). f , RNA-seq analysis of CD62L + TCR + LN T cells from CD8 Dual and B6 mice. Genes differentially expressed between B6 CD4 + and CD8 + LN T cells were evaluated in the heat map for analysis of gene expression in CD8.1 and CD8.2 LN T cells from CD8 Dual mice ( n = 3 per group, P < 0.05, 5-fold change). Numbers within profiles and histograms indicate frequency of cells in each box or lines ( b and c ). *** P < 0.001, ** P < 0.01, * P < 0.05 (two-tailed unpaired t -tests); mean ± s.e.m. ( d and e ).
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    strain b6 129p2 β2m tm1 unc j  (Jackson Laboratory)
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    Jackson Laboratory strain b6 129p2 β2m tm1 unc j
    a , Top: schematic of double-positive thymocytes from CD8 Dual mice. The altered Cd4 gene locus encodes CD8α.1 co-receptors (composed of CD8α.1CD8β dimers), and the endogenous Cd8 gene locus encodes CD8α.2 co-receptors (composed of CD8α.2CD8β dimers). Bottom: schematic of the altered Cd4 CD8 and endogenous Cd8 CD8 genes in CD8 Dual mice; IRES, internal ribosome entry site; poly A, polyadenylation signals. b , Flow cytometry analysis of whole thymocytes from CD8 Dual ( n = 16) and B6 ( n = 15) mice (representative of 15 independent experiments). Top: Cd4 -encoded co-receptors (CD8α.1 or CD4) versus Cd8 -encoded co-receptor CD8α.2 profile, showing four thymocyte subsets; double-negative (DN), double-positive (DP) and two single-positive T cell subsets. Total cell number (mean ± s.e.m) is shown above profiles. Middle: TCRβ histogram, showing TCRβ hi (TCR hi ) thymocytes. Bottom: CD24 versus TCRβ profile, showing CD24 − TCR + mature thymocytes. c , Flow cytometry analysis of TCR hi thymocytes and TCR + LN T cells from CD8 Dual ( n = 16) and B6 ( n = 15) mice, showing Cd4 -encoded co-receptor (CD8α.1 or CD4) versus Cd8 -encoded co-receptor (CD8α.2) or CD8β profiles. TCR expression of LN cells are shown in Extended Data Fig. . Numbers (mean ± s.e.m.) of TCR hi thymocytes and LN T cells are shown above profiles (representative of 15 independent experiments). d , Numbers of CD8β + T cells among CD24 − TCR + mature thymocytes (T) and TCR + LN T cells (L) in CD8 Dual mice with the indicated MHC deficiencies. CD8β histograms are shown in Extended Data Fig. (WT: T ( n = 6), L ( n = 9), MHC-II KO : T ( n = 9), L ( n = 9), <t>β2m</t> KO : T ( n = 7), L ( n = 9), MHC KO : T ( n = 3), L ( n = 5), 3–7 independent experiments). e , Mean fluorescence intensity (MFI) of Runx3d-YFP and ThPOK-GFP reporter expression in CD8.1, CD8.2, CD4 + and CD8 + T cells among TCR hi thymocytes and TCR + LN T cells in CD8 Dual and B6 mice based on histograms shown in Extended Data Fig. (Runx3d-YFP: CD8 Dual T ( n = 4), L ( n = 4), B6 T ( n = 5), L ( n = 4), ThPOK-GFP: CD8 Dual T ( n = 3), L ( n = 3), B6 T ( n = 4) L ( n = 3), representative of 3–5 independent experiments). f , RNA-seq analysis of CD62L + TCR + LN T cells from CD8 Dual and B6 mice. Genes differentially expressed between B6 CD4 + and CD8 + LN T cells were evaluated in the heat map for analysis of gene expression in CD8.1 and CD8.2 LN T cells from CD8 Dual mice ( n = 3 per group, P < 0.05, 5-fold change). Numbers within profiles and histograms indicate frequency of cells in each box or lines ( b and c ). *** P < 0.001, ** P < 0.01, * P < 0.05 (two-tailed unpaired t -tests); mean ± s.e.m. ( d and e ).
    Strain B6 129p2 β2m Tm1 Unc J, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a , Top: schematic of double-positive thymocytes from CD8 Dual mice. The altered Cd4 gene locus encodes CD8α.1 co-receptors (composed of CD8α.1CD8β dimers), and the endogenous Cd8 gene locus encodes CD8α.2 co-receptors (composed of CD8α.2CD8β dimers). Bottom: schematic of the altered Cd4 CD8 and endogenous Cd8 CD8 genes in CD8 Dual mice; IRES, internal ribosome entry site; poly A, polyadenylation signals. b , Flow cytometry analysis of whole thymocytes from CD8 Dual ( n = 16) and B6 ( n = 15) mice (representative of 15 independent experiments). Top: Cd4 -encoded co-receptors (CD8α.1 or CD4) versus Cd8 -encoded co-receptor CD8α.2 profile, showing four thymocyte subsets; double-negative (DN), double-positive (DP) and two single-positive T cell subsets. Total cell number (mean ± s.e.m) is shown above profiles. Middle: TCRβ histogram, showing TCRβ hi (TCR hi ) thymocytes. Bottom: CD24 versus TCRβ profile, showing CD24 − TCR + mature thymocytes. c , Flow cytometry analysis of TCR hi thymocytes and TCR + LN T cells from CD8 Dual ( n = 16) and B6 ( n = 15) mice, showing Cd4 -encoded co-receptor (CD8α.1 or CD4) versus Cd8 -encoded co-receptor (CD8α.2) or CD8β profiles. TCR expression of LN cells are shown in Extended Data Fig. . Numbers (mean ± s.e.m.) of TCR hi thymocytes and LN T cells are shown above profiles (representative of 15 independent experiments). d , Numbers of CD8β + T cells among CD24 − TCR + mature thymocytes (T) and TCR + LN T cells (L) in CD8 Dual mice with the indicated MHC deficiencies. CD8β histograms are shown in Extended Data Fig. (WT: T ( n = 6), L ( n = 9), MHC-II KO : T ( n = 9), L ( n = 9), β2m KO : T ( n = 7), L ( n = 9), MHC KO : T ( n = 3), L ( n = 5), 3–7 independent experiments). e , Mean fluorescence intensity (MFI) of Runx3d-YFP and ThPOK-GFP reporter expression in CD8.1, CD8.2, CD4 + and CD8 + T cells among TCR hi thymocytes and TCR + LN T cells in CD8 Dual and B6 mice based on histograms shown in Extended Data Fig. (Runx3d-YFP: CD8 Dual T ( n = 4), L ( n = 4), B6 T ( n = 5), L ( n = 4), ThPOK-GFP: CD8 Dual T ( n = 3), L ( n = 3), B6 T ( n = 4) L ( n = 3), representative of 3–5 independent experiments). f , RNA-seq analysis of CD62L + TCR + LN T cells from CD8 Dual and B6 mice. Genes differentially expressed between B6 CD4 + and CD8 + LN T cells were evaluated in the heat map for analysis of gene expression in CD8.1 and CD8.2 LN T cells from CD8 Dual mice ( n = 3 per group, P < 0.05, 5-fold change). Numbers within profiles and histograms indicate frequency of cells in each box or lines ( b and c ). *** P < 0.001, ** P < 0.01, * P < 0.05 (two-tailed unpaired t -tests); mean ± s.e.m. ( d and e ).

    Journal: Nature Immunology

    Article Title: Unraveling CD8 lineage decisions reveals that functionally distinct CD8 + T cells are selected by different MHC-I thymic peptides

    doi: 10.1038/s41590-025-02411-4

    Figure Lengend Snippet: a , Top: schematic of double-positive thymocytes from CD8 Dual mice. The altered Cd4 gene locus encodes CD8α.1 co-receptors (composed of CD8α.1CD8β dimers), and the endogenous Cd8 gene locus encodes CD8α.2 co-receptors (composed of CD8α.2CD8β dimers). Bottom: schematic of the altered Cd4 CD8 and endogenous Cd8 CD8 genes in CD8 Dual mice; IRES, internal ribosome entry site; poly A, polyadenylation signals. b , Flow cytometry analysis of whole thymocytes from CD8 Dual ( n = 16) and B6 ( n = 15) mice (representative of 15 independent experiments). Top: Cd4 -encoded co-receptors (CD8α.1 or CD4) versus Cd8 -encoded co-receptor CD8α.2 profile, showing four thymocyte subsets; double-negative (DN), double-positive (DP) and two single-positive T cell subsets. Total cell number (mean ± s.e.m) is shown above profiles. Middle: TCRβ histogram, showing TCRβ hi (TCR hi ) thymocytes. Bottom: CD24 versus TCRβ profile, showing CD24 − TCR + mature thymocytes. c , Flow cytometry analysis of TCR hi thymocytes and TCR + LN T cells from CD8 Dual ( n = 16) and B6 ( n = 15) mice, showing Cd4 -encoded co-receptor (CD8α.1 or CD4) versus Cd8 -encoded co-receptor (CD8α.2) or CD8β profiles. TCR expression of LN cells are shown in Extended Data Fig. . Numbers (mean ± s.e.m.) of TCR hi thymocytes and LN T cells are shown above profiles (representative of 15 independent experiments). d , Numbers of CD8β + T cells among CD24 − TCR + mature thymocytes (T) and TCR + LN T cells (L) in CD8 Dual mice with the indicated MHC deficiencies. CD8β histograms are shown in Extended Data Fig. (WT: T ( n = 6), L ( n = 9), MHC-II KO : T ( n = 9), L ( n = 9), β2m KO : T ( n = 7), L ( n = 9), MHC KO : T ( n = 3), L ( n = 5), 3–7 independent experiments). e , Mean fluorescence intensity (MFI) of Runx3d-YFP and ThPOK-GFP reporter expression in CD8.1, CD8.2, CD4 + and CD8 + T cells among TCR hi thymocytes and TCR + LN T cells in CD8 Dual and B6 mice based on histograms shown in Extended Data Fig. (Runx3d-YFP: CD8 Dual T ( n = 4), L ( n = 4), B6 T ( n = 5), L ( n = 4), ThPOK-GFP: CD8 Dual T ( n = 3), L ( n = 3), B6 T ( n = 4) L ( n = 3), representative of 3–5 independent experiments). f , RNA-seq analysis of CD62L + TCR + LN T cells from CD8 Dual and B6 mice. Genes differentially expressed between B6 CD4 + and CD8 + LN T cells were evaluated in the heat map for analysis of gene expression in CD8.1 and CD8.2 LN T cells from CD8 Dual mice ( n = 3 per group, P < 0.05, 5-fold change). Numbers within profiles and histograms indicate frequency of cells in each box or lines ( b and c ). *** P < 0.001, ** P < 0.01, * P < 0.05 (two-tailed unpaired t -tests); mean ± s.e.m. ( d and e ).

    Article Snippet: Aire KO (ref. ), BALB/cJ, β2m KO (ref. ), CD1d KO (ref. ), CD8α KO (ref. ), IL-4R KO (ref. ) and IL-15 KO (ref. ) mice were purchased from The Jackson Laboratory. β5t KO from Y.T., PLZF KO from D. Kovalovsky , MHC-II KO , HY.Rag2 KO , P14.Rag2 KO , OT-I.Rag2 KO , Rag-GFP transgene (Tg) from M. Nussenzweig , Runx3d-YFP knock-in from D. R. Littman and ThPOK-GFP Tg mice from R. Bosselut were maintained in our own animal colony.

    Techniques: Flow Cytometry, Expressing, Fluorescence, RNA Sequencing, Gene Expression, Two Tailed Test