MR1 抗体 (AA 201-300)
(3 validations)-
- 抗原 See all MR1 抗体
- MR1 (Major Histocompatibility Complex, Class I-Related (MR1))
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抗原表位
- AA 201-300
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适用
- 人
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宿主
- 小鼠
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克隆类型
- 单克隆
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标记
- This MR1 antibody is un-conjugated
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应用范围
- ELISA, Western Blotting (WB), Immunofluorescence (IF)
- 原理
- Mouse monoclonal antibody raised against a partial recombinant MR1.
- 序列
- TEPPLVRVNR KETFPGVTAL FCKAHGFYPP EIYMTWMKNG EEIVQEIDYG DILPSGDGTY QAWASIELDP QSSNLYSCHV EHCGVHMVLQ VPQESETIPL
- 交叉反应
- 人
- 产品特性
- Antibody Reactive Against Recombinant Protein.
- 免疫原
- MR1 (NP_001522, 201 a.a. ~ 300 a.a) partial recombinant protein with GST tag. MW of the GST tag alone is 26 KDa.
- 克隆位点
- 5B5
- 亚型
- IgG1
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anti-Major Histocompatibility Complex, Class I-Related (MR1) (AA 312-341), (C-Term) antibody
MR1 适用: 人 WB 宿主: 兔 Polyclonal RB37199 unconjugated
anti-Major Histocompatibility Complex, Class I-Related (MR1) (AA 150-253) antibodyMR1 适用: 人 ELISA, WB, IHC, IF 宿主: 兔 Polyclonal unconjugated
anti-Major Histocompatibility Complex, Class I-Related (MR1) (AA 20-260) antibodyMR1 适用: 人 WB, IF 宿主: 兔 Polyclonal unconjugated
anti-Major Histocompatibility Complex, Class I-Related (MR1) (AA 150-253) antibodyMR1 适用: 人, 小鼠 ELISA, WB, IHC, IF/ICC 宿主: 兔 Polyclonal unconjugated
anti-Major Histocompatibility Complex, Class I-Related (MR1) (AA 20-260) antibodyMR1 适用: 人 ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-Major Histocompatibility Complex, Class I-Related (MR1) (AA 201-300) antibodyMR1 适用: 人 ELISA, WB 宿主: 小鼠 Polyclonal unconjugated
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- 应用备注
- Optimal working dilution should be determined by the investigator.
- 限制
- 仅限研究用
-
- by
- Dr. Randy Brutkiewicz Laboratory, Department of Microbiology and Immunology, Indiana University School of Medicine
- No.
- #101752
- 日期
- 2018.02.20
- 抗原
- MR1
- Lot Number
- 12045-5B5
- Method validated
- Immunocytochemistry
- Positive Control
- HEK293 cells transfected with human MR1 cDNA
- Negative Control
- HEK293 cells transfected with plasmid vector only
- Notes
Passed. The MR1 antibody ABIN516526 specifically labels the targeted antigen in HEK293 ectopically expressing human MR1 in ICC.
- Primary Antibody
- ABIN516526
- Secondary Antibody
- FITC-conjugated donkey anti-mouse immunoglobulin antiserum (Jackson ImmunoResearch, 715-096-151, lot 76750)
- Full Protocol
- Grow HEK293 cells in DMEM medium (Lonza, 12-614F, lot 0000618582) supplemented with serum (Hyclone, SH30071.03, lot AAG205460) and antibiotics (Hyclone, SV30010, lot J150013), at 37°C and 5% CO2 dish to 70-90% confluency.
- Transfect cells with pCDNA 3.1 neo (-) (Invitrogen) containing human MR1 cDNA (Genecopoeia) using Polyethylenimine (Polysciences, 23966) following the manufacturer´s instructions.
- Plate human MR1-expressing HEK293 cells in sterile glass-bottom 35mm dishes coated with collagen (MatTek, P35GCol-1.5-14-C) to 50-80% confluency.
- Wash cells PBS.
- Fix cells with 4% Paraformaldehyde for 15min at RT.
- Block cells with blocking buffer (1x PBS, 5% donkey serum, 0.3% Triton X-100) for 1h at RT.
- Incubate cells with primary mouse anti-MR1 antibody (antibodies-online, ABIN516526, lot 12045-5B5) diluted 1:50 in dilution buffer (1x PBS, 1% BSA, 0.3% Triton X-100) and incubated ON at 4°C.
- Wash cells 3x with PBS.
- Incubated with secondary FITC-conjugated donkey anti-mouse immunoglobulin antiserum (Jackson ImmunoResearch, 715-096-151, lot 76750) diluted 1:50 in dilution buffer for 1h at RT.
- Wash cells 3x with PBS.
- To stain the nucleus, immerse cells in PBS containing Hoechst (1:2000) for 5min.
- Just prior to confocal analysis, place cells in mounting medium (10mM Tris pH8.5, 2% DABCO).
- Image cells on an Olympus 2 confocal/two-photon microscope imaging system using an oil immersion lens at 60×.
- Experimental Notes
Staining with ABIN516526 shows a perinuclear pattern, suggesting MR1 localizes in the endoplasmic reticulum. No signal was detected in sample negative control tissue and the secondary antibody only control.
生效 #101752 (Immunocytochemistry)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Human MR1-expressing HEK293 cells were stained with MR1 antibody ABIN516526 and a FITC-conjugated secondary antibody (green, A). For nuclear staining, cells were stained with Hoechst (blue, B). C shows both channels merged.]()
Human MR1-expressing HEK293 cells were stained with MR1 antibody ABIN516526 and a FITC-conjugated secondary antibody (green, A). For nuclear staining, cells were stained with Hoechst (blue, B). C shows both channels merged.
Full Methods -
- by
- Dr. Randy Brutkiewicz Laboratory, Department of Microbiology and Immunology, Indiana University School of Medicine
- No.
- #102826
- 日期
- 2018.02.20
- 抗原
- MR1
- Lot Number
- 12045-5B5
- Method validated
- Immunoprecipitation
- Positive Control
- HEK293 cells transfected with human MR1 cDNA
- Negative Control
- HEK293 cells transfected with plasmid vector only
- Notes
Passed. ABIN516526 immunoprecipitates human MR1 overexpressed by HEK293 cells.
- Primary Antibody
- ABIN516526
- Secondary Antibody
- goat anti-rabbit Dye-IR800 conjugated antibody (Advansta, R-05060-250, lot 17083179)
- Full Protocol
- Grow HEK293 cells in DMEM medium (Lonza, 12-614F, lot 0000618582) supplemented with serum (Hyclone, SH30071.03, lot AAG205460) and antibiotics (Hyclone, SV30010, lot J150013), at 37°C and 5% CO2 dish to 70-90% confluency.
- Transfect cells with pCDNA 3.1 neo (-) (Invitrogen) containing human MR1 cDNA (Genecopoeia) using Polyethylenimine (Polysciences, 23966) following the manufacturer´s instructions.
- Lyse cells in cold lysis buffer (10mM Tris pH7.4, 150mM NaCl, 0.5mM EDTA, 2% CHAPS).
- Determine total protein content of the lysates using Commassie Protein Assay Reagent (Thermo Scientific, 1856209, lot NL179252).
- Immobilize 100µl of protein G-conjugated Sepharose beads (Pierce, product 20399, lot RI239318) ON at 4°C with
- 2.5µg mouse anti-MR1 antibody (antibodies-online, ABIN2665876, lot B177559),
- 2.5µg mouse anti-MR1 antibody (antibodies-online, ABIN516526, lot12045-5B5),
- 2.5µg rabbit anti-MR1 antibody (antibodies-online, ABIN1537116, lot SA111213CH),
- 2.5µg mouse IgG2a antibody (Biolegend, 400202, lot B153642),
- 2.5µg mouse IgG1 antibody (BD, 555746, lot 3221830), or
- 2.5µg rabbit IgG antibody (Santa Cruz Biotechnology, SC-5560, lot E0609).
- Incubate 500µg of the cell lysates with 2.5µg of antibody-bead conjugate ON at 4°C.
- Wash lysates 4x with PBS.
- Denature beads for 5min at 95°C in 60µl Laemmli SDS sample buffer and subsequently separate them on a SDS-PAGE gel using Acrylamide/Bis Premixed (Bio-Rad, 61-0125, lot 260000477) for 2-3h at 100V.
- Transfer proteins onto PVDF membrane (Millipore, IPVH00010, lot K5AA6843U) with a Western blotting system for ON at 4°C at 150mA.
- Block the membrane with blocking buffer (2% BSA/PBS/0.05%Tween-20) for 1h at RT.
- Incubate membrane with primary rabbit anti-MR1 antibody (antibodies-online ABIN1537116, lot SA111213CH) diluted 1:1000 in blocking buffer ON at 4°C.
- Wash membrane 3x for 10min with PBS/0.05%Tween-20.
- Incubate membrane with secondary goat anti-rabbit Dye-IR800 conjugated antibody (Advansta, R-05060-250, lot 17083179) diluted 1:10000 in PBS/0.05% Tween-20 for 1h at RT.
- Wash membrane 3x for 10 min with PBS/0.05% Tween-20.
- Reveal protein bands using an Odyssey imaging system (LI-COR Biosciences).
- Experimental Notes
The human MR1 antibody ABIN516526, but not the isotype control, immunoprecipitates with human MR1 overexpressed by HEK293 cells.
生效 #102826 (Immunoprecipitation)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Lysates from human MR1-expressing HEK293 cells were immunoprecipitated by antibodies specific for MR1 (ABIN2665876, ABIN516526, ABIN1537116) or the respective isotype controls (mouse IgG2a, mouse, IgG1, rabbit IgG). Immunoprecipitants were resolved by SDS-PAGE gel followed by Western blotting analysis using MR1 antibody ABIN1537116.]()
Lysates from human MR1-expressing HEK293 cells were immunoprecipitated by antibodies specific for MR1 (ABIN2665876, ABIN516526, ABIN1537116) or the respective isotype controls (mouse IgG2a, mouse, IgG1, rabbit IgG). Immunoprecipitants were resolved by SDS-PAGE gel followed by Western blotting analysis using MR1 antibody ABIN1537116.
Full Methods -
- by
- Dr. Randy Brutkiewicz Laboratory, Department of Microbiology and Immunology, Indiana University School of Medicine
- No.
- #102827
- 日期
- 2018.02.20
- 抗原
- MR1
- Lot Number
- 12045-5B5
- Method validated
- Flow Cytometry
- Positive Control
- HEK293 cells transfected with human MR1 cDNA
- Negative Control
- HEK293 cells transfected with plasmid vector only
- Notes
Passed. ABIN2665876 recognizes human MR1 overexpressed by HEK293 cells and can be used in flow cytometry.
- Primary Antibody
- ABIN516526
- Secondary Antibody
- PE-conjugated rabbit anti-mouse immunoglobulin antiserum (Jackson ImmunoResearch, 115-116-146, lot 120701)
- Full Protocol
- Grow HEK293 cells in DMEM medium (Lonza, 12-614F, lot 0000618582) supplemented with serum (Hyclone, SH30071.03, lot AAG205460) and antibiotics (Hyclone, SV30010, lot J150013), at 37°C and 5% CO2 dish to 70-90% confluency.
- Transfect cells with pCDNA 3.1 neo (-) (Invitrogen) containing human MR1 cDNA (Genecopoeia) using Polyethylenimine (Polysciences, 23966) following the manufacturer´s instructions.
- Surface staining:
- Wash 0.5x106 cells 3x with HBSS/0.1% BSA.
- Incubate cells with mouse anti-MR1 antibody (antibodies-online, ABIN516526, lot 12045-5B5) diluted 1:100 or mouse anti-MR1 antibody (antibodies-online, ABIN2665876, lot B177559) diluted 1:100 for 30min on ice.
- Wash cells 3x with HBSS/0.1% BSA.
- Incubate cells with a PE-conjugated rabbit anti-mouse immunoglobulin antiserum (Jackson ImmunoResearch, 115-116-146, lot 120701) diluted 1:100 for 30min on ice.
- Total cell staining:
- Fix 0.5x106 cells in 1% paraformaldehyde for 10min at RT.
- Wash cells with HBSS/BSA.
- Permeabilize cells in HBSS/BSA with 0.1% saponin for 10min at RT.
- Incubate cells with mouse anti-MR1 antibody (antibodies-online, ABIN516526, lot 12045-5B5) diluted 1:100 or mouse anti-MR1 antibody (antibodies-online, ABIN2665876, lot B177559) dilute 1:100 for 30min at RT in the presence of 0.1% saponin.
- Wash cells 3x with HBSS/0.1% BSA/0.1% saponin.
- Incubate cells with PE-conjugated rabbit anti-mouse immunoglobulin antiserum (Jackson ImmunoResearch, 115-116-146, lot 120701) diluted 1:100 for 30min at RT.
- Acquire flow cytometry data using a flow cytometry using LSR 4 (BD Biosciences) followed data analysis using FlowJo software.
- Experimental Notes
The human MR1 antibody ABIN2665876 shows specific staining for human MR1.
生效 #102827 (Flow Cytometry)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Human MR1-expressing HEK293 cells (MR1) and vector control cells (Vc) were stained with MR1 antibodies ABIN516526 or ABIN2665876 followed by PE-conjugated anti-mouse secondary antibody. For total MR1 staining, cells were permeabilized with 0.1% saponin and stained with primary and secondary antibodies. Cells were analyzed by flow cytometry.]()
Human MR1-expressing HEK293 cells (MR1) and vector control cells (Vc) were stained with MR1 antibodies ABIN516526 or ABIN2665876 followed by PE-conjugated anti-mouse secondary antibody. For total MR1 staining, cells were permeabilized with 0.1% saponin and stained with primary and secondary antibodies. Cells were analyzed by flow cytometry.
Full Methods -
- 缓冲液
- In 1x PBS, pH 7.4
- 注意事项
- Aliquot to avoid repeated freezing and thawing.
- 储存条件
- -20 °C
- 储存方法
- Store at -20°C or lower. Aliquot to avoid repeated freezing and thawing.
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- 抗原
- MR1 (Major Histocompatibility Complex, Class I-Related (MR1))
- 别名
- MR1 (MR1 产品)
- 别名
- YFV antibody, hlals antibody, MGC154362 antibody, MR1 antibody, DKFZp468C1823 antibody, HLALS antibody, H2ls antibody, Hlals antibody, MHC class I antigen YF5 antibody, major histocompatibility complex, class I-related antibody, major histocompatibility complex, class I-related L homeolog antibody, YF5 antibody, MR1 antibody, mr1.L antibody, mr1 antibody, Mr1 antibody
- 背景
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Full Gene Name: major histocompatibility complex, class I-related
Synonyms: HLALS - 基因ID
- 3140
- NCBI登录号
- NM_001531
- 途径
- Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Cancer Immune Checkpoints
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