Invitrogen

MHC Class I (H-2Kb) Monoclonal Antibody (AF6-88.5.5.3), Alexa Fluor™ 700, eBioscience™

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MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

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MHC Class I (H-2Kb) Antibody (56-5958-82) in Flow

Staining of BALB/c (left) and C57BL/6 (right) splenocytes with 0.25 µg of Mouse IgG2a kappa Isotype Control, Alexa Fluor 700 (Product # 56-4724-82) (blue histogram) or 0.25 µg of MHC Class I (H-2Kb) Monoclonal Antibody, Alexa Fluor 700 (purple histogram). Total cells were used for analysis.

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow)

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MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Staining of BALB/c (left) and C57BL/6 (right) splenocytes with 0.25 µg of Mouse IgG2a kappa Isotype Control, Alexa Fluor 700 (Product # 56-4724-82) (blue histogram) or 0.25 µg of MHC Class I (H-2Kb) Monoclonal Antibody, Alexa Fluor 700 (purple histogram). Total cells were used for analysis.

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Figure 6 AAP inhibits 4T1 tumor growth in vivo. ( A ) BALB/c mice containing 4T1 tumors were treated 2x/week with NAC (100 mg/kg)/propylene glycol (PG, 10%) or AAP (500 mg/kg)/NAC/PG and tumor growth was monitored with digital calipers. ( B ) At time of animal sacrifice, tumors were weighed. ( C ) At conclusion of study, tumors were digested and protein levels of IFN gamma and IL-10 determined using ELISA. ( D ) CD45+/CD11B+/F4/80+ macrophages were analyzed for M1 and M2 macrophage markers by flow cytometry. ( E ) Average MFI of indicated markers in macrophages from 3 mice tumors per treatment group are shown. N = 3 per condition. * <=0.05. Error bars represent +-1 SD.

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Figure 5 AAP inhibits EF43.fgf4 tumor growth in vivo. ( A ) BALB/c mice containing EF43.fgf4 (EF43) tumors were treated 2x/week with NAC (100 mg/kg)/Propylene glycol (PG, 10%) or AAP (500 mg/kg)/NAC/PG and tumor growth monitored with digital calipers. ( B ) At time of animal sacrifice, tumors were weighed. ( C ) At conclusion of study, tumors were digested and analyzed for IL-10 and IFN gamma by ELISA. ( D ) CD45+/CD11B+/F4/80+ macrophages were analyzed for M1 and M2 macrophage markers by flow cytometry. ( E ) Average MFI of indicated markers in macrophages from 3 mice tumors per treatment group are shown. * <=0.05. Error bars represent +-1 SD.

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Characterization of mouse myeloid-DC (Mono-DC) and hematopoietic stem-DC (Stem-DC) derived from BM cells pulsed with tumor antigens of ovarian cancer. Mono-DC differentiation is shown from ( A-D ), and stem-DC differentiation is shown from ( E-G ). ( A ) Images of differentiated Mono-DCs at 7 days after BM cell cultures were treated with GM-CSF and IL-4 and pulsed with tumor cell lysates. ( B ) Flow cytometry analysis of differentiated Mono-DCs showing the expression of DC-specific cell surface markers and the absence of CD8alpha and Clec9a, which are intrinsic characteristics of mouse Stem-DCs. ( C ) Cytokine secretion by differentiated Mono-DCs with or without maturation factors was tested by ELISA at 24 h after stimulation (P = 0.0445, < 0.0001, and 0.0001, respectively). ( D ) T cell proliferation (MLR assay) with different amounts of Mono-DCs (2:1, DC 5 x 10 4 cells + /- T cell 1 x 10 5 cells; 5:1, DC 2 x 10 4 cells + /- T cell 1 x 10 5 cells; 10:1, DC 1 x 10 4 cells + /- T cell 1 x 10 5 cells; T cell only, T cell 1 x 10 5 cells) (P = 0.0030 and P = 0.0044, respectively). ( E ) The typical phenotype of mouse Stem-DC (CD8alpha, Clec9a) was confirmed by FACS analysis. ( F ) IL-12 and IFN-gamma secretion by differentiated stem-DCs with or without maturation factors was tested by ELISA immediately after culture (P = 0.0143 and P = 0.0286, respectively). ( G ) T cell proliferation (MLR assay) with stem-DCs in the presence or absence of maturation factors showed a significant

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Fig. 6 MiR-223 reduced donor T cell migration to recipient mice spleens. The proportion of H2kb + CD4 and CD8 T cell in recipient mice spleens on day 7 was assessed by flow cytometry. Compared with NC and aGvHD group, H2kb + CD4 and H2kb + CD8 cell percentages in miR-223 group showed significant reduction ( a ). The average number of H2kb + CD4 and H2kb + CD8 cells in miR-223 group showed significant decline ( b ). Data were presented as mean +- SEM. Measured in three independent experiments, n = 7 per group. * P < 0.05, ** P < 0.01

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Fig. 3 Tumor cells lysed by M1 mature iDCs and enable the iDCs to prime de novo T-cell responses ex vivo. A Diagrammatic sketch of the coculture experiment performed ex vivo. B16F10 cells were treated with ctrl, M1, UV-inactivated M1, or physical lysis on day 0, and then the supernatant of each group was collected and fed to immature DCs (iDCs) on day 2. One batch of DCs was used to detect maturation markers by flow cytometry and secreted cytokines by ELISA on day 4. The other batch of DCs was cocultured with naive T cells on day 4, and T-cell activation was evaluated on day 8. B , C MHC I ( B ) and CD86 ( C ) expression on DCs fed the indicated supernatants was detected by flow cytometry on day 4. n = 3. P values were determined by one-way ANOVA with Dunnett's tests for multiple comparisons. D IL-12, TNF-alpha, and IL-1beta levels in the culture medium of DCs fed with the indicated supernatants were determined by ELISA on day 4. n = 3. The P value was determined by a two-tailed Student's t test. E Phase-contrast images of proliferating T-cell clones on day 8 are shown. The images are representative of three independent experiments. Scale bars, 50 mum. F IFN-gamma, granzyme B, and perforin concentrations were determined by ELISA in the DC plus T-cell coculture system on day 8. n = 3. The P value was determined by a two-tailed Student's t test. G-H, CD69, and CD44 expression on CD4 + T cells ( G ) and CD8 + T cells ( H ) was evaluated on T cells in the DC plus T-cell coculture

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Figure 1 A lack of platelet-derived beta2M changes age associated monocyte phenotypes. ( A ) Platelets are a major source of age associated increases in plasma beta2M. Plasma was isolated from 2-4 and 14-16 mos old WT and Plt-beta2M -/- mice and beta2M was quantified by ELISA. Plasma beta2M had a greater increase with age in WT compared to Plt-beta2M -/- mice (N=11, mean +- SEM, *P<0.05, **P<0.01, one-way ANOVA with Bonferroni correction). ( B ) Platelet MHC I increased with age. Platelets were isolated from 4 and 14 mos old WT and Plt-beta2M -/- mice. Surface MHC I was quantified by flow cytometry. MHC I increased with age in WT, but not Plt-beta2M -/- mice (N=3-4, mean +- SEM, **P<0.01, one-way ANOVA with Bonferroni correction). ( C ) Plasma TGFbeta does not significantly change with age in either WT or Plt-beta2M -/- mice. Plasma TGFbeta was quantified by ELISA (N=17-24, mean +- SEM, one-way ANOVA with Bonferroni correction). ( D ) Aged Plt-beta2M -/- mice had fewer circulating Ly6C Hi monocytes compared to WT mice. Peripheral blood was isolated from 4 mos and 14 mos old WT and Plt-beta2M -/- mice and monocyte Ly6C expression determined by flow cytometry. Ly6C Hi monocytes were increased in older WT, but not Plt-beta2M -/- mice (N=8-31, mean +- SEM, **P<0.01, one-way ANOVA with Bonferroni correction). ( E ) 14 mos old WT mice have increased plasma KC while Plt-beta2M -/- have increased plasma IL-10 (N=16-22, mean +- SEM, one-way ANOVA with Bonferroni correction). ( F ) 14

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Figure 6. CXCL14-mediated tumor suppression is abrogated by MHC-I knockdown in HNC cells. MOE/E6E7 CXCL14 cells were transduced with shRNAs directed to B2M and enriched for knockdown by FACS sorting for the bottom 10% of MHC-I expressing cells (MOE/E6E7 CXCL14/shB2M ). Flow cytometric analysis of H-2D b (A) and H-2K b (B) MHC-I molecules in MOE/E6E7 Vector (gray), MOE/E6E7 CXCL14 (blue), and MOE/E6E7 CXCL14/shB2M (red) cells by flow cytometry. Wildtype B6 mice ( n = 10 per group) were s.c. injected with MOE/E6E7 Vector , MOE/E6E7 CXCL14 , or MOE/E6E7 CXCL14/shB2M cells (5 x 10 5 cells/mouse). Tumor volume was measured twice per week (C). Individual growth curves are shown from mice injected with MOE/E6E7 CXCL14 (E) and MOE/E6E7 CXCL14/shB2M (F) cells. Survival rates of mice injected with MOE/E6E7 Vector , MOE/E6E7 CXCL14 , and MOE/E6E7 CXCL14/shB2M cells were analyzed as was performed in Fig. 1 (D). Summary model of findings of CXCL14 on peripheral immune cells and on immune gene expression (G) P value of MOE/E6E7 CXCL14/shB2M cells compared to MOE/E6E7 Vector or MOE/E6E7 CXCL14 cells was determined for tumor growth (C) and survival (D) by two-way ANOVA analysis. represents mice that exhibited tumor growth but had to be euthanized due to self-inflicted wounds. * p < 0.01, ** p < 0.001. Shown are representative of two independent experiments.

MHC Class I (H-2Kb) Antibody in Flow Cytometry (Flow) — Figure 5. CXCL14 expression restores surface expression of MHC-I proteins on HPV-positive HNC cells. Surface expression of MHC-I haplotype proteins, H-2D b (A-C) and H-2K b (D and E), was determined in normal immortalized MOE (NI-MOE) cells (purple) (A), parental MOE/E6E7 cells (gray) (A), MOE/E6E7 Vector cells (gray) (B), and two clones of MOE/E6E7 CXCL14 cells (CL8-red and CL16-blue) (B) by flow cytometry. Isotype controls were evaluated on NI-MOE cells (purple outline with light purple fill) and parental MOE/E6E7 cells (black outline). Relative mean fluorescent intensity (MFI) values of MOE/E6E7 Vector and MOE/E6E7 CXCL14-CL16 cells were calculated (C). P value was determined by the Student's t -test. * p < 0.05. IFNgamma treatment was used as a positive control to validate H-2K b expression in MOE/E6E7 CXCL14 cells (E). Shown are representative of three independent experiments.

产品信息

56-5958-82

应用

建议稀释比

已发表文章

流式细胞分析 (Flow)

0.5 µg/test

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产品规格

种属反应

Mouse

已发表种属

Not Applicable

宿主/亚型

Mouse / IgG2a, kappa

推荐的同型对照

Mouse IgG2a kappa Isotype Control (eBM2a), Alexa Fluor™ 700, eBioscience™

分类

Monoclonal

类型

Antibody

克隆号

AF6-88.5.5.3

偶联物

Alexa Fluor™ 700 Alexa Fluor™ 700 Alexa Fluor™ 700

激发/发射光谱

696/719 nm 查看光谱

形式

Liquid

浓度

0.2 mg/mL

纯化类型

Affinity chromatography

保存液

PBS, pH 7.2

内含物

0.09% sodium azide

保存条件

4° C, store in dark, DO NOT FREEZE!

运输条件

Ambient (domestic); Wet ice (international)

AB_2815254

产品详细信息

Description: This AF6-88.5.5.3 monoclonal antibody reacts with the H-2Kb MHC class I alloantigen. H-2Kb is involved in antigen presentation to T cells expressing CD3/TCR and CD8. Reactivity to other haplotypes (e.g. d, f, j, k, p, q, r, s, u, and v) has not been observed.

Applications Reported: This AF6-88.5.5.3 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This AF6-88.5.5.3 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 700 emits at 723 nm and can be excited with the red laser (633-647 nm). Most instruments will require a 685 LP mirror and 710/20 band pass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

Excitation: 633-647 nm; Emission: 723 nm; Laser: Red Laser

靶标信息

This protein is an MHC class I alloantigen, which is involved in antigen presentation to T cells expressing CD3/TCR and CD8.

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.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

How to use the Panel Builder

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生物信息学

蛋白别名: class I major histocompatibility antigen H-2Kd; H-2 class I histocompatibility antigen, K-B alpha chain; H-2 class I histocompatibility antigen, K-D alpha chain; H-2 class I histocompatibility antigen, K-K alpha chain; H-2 class I histocompatibility antigen, K-Q alpha chain; H-2 class I histocompatibility antigen, K-W28 alpha chain; H-2K(B); H-2K(K); H-2K(Q); MHC class I antigen; MHC class I H2-K-b-alpha-2 cell surface glycoprotein; MHC class I heavy chain H2-K; MHC H2-K transplantation antigen

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基因别名: H-2K; H-2K(d); H2-K; H2-K1; K-f

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UniProt ID: (Mouse) P01901

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Entrez Gene ID: (Mouse) 14972

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receptor binding protein binding beta-2-microglobulin binding peptide binding peptide antigen binding T cell receptor binding poly(A) RNA binding TAP binding protein heterodimerization activity major histocompatibility complex protein

positive regulation of T cell mediated cytotoxicity immune system process antigen processing and presentation of peptide antigen via MHC class I antigen processing and presentation of endogenous peptide antigen via MHC class I via ER pathway, TAP-dependent immune response antigen processing and presentation antigen processing and presentation of exogenous peptide antigen via MHC class I defense response to bacterium inner ear development