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MR1 抗体 (C-Term)

MR1 适用: 人 WB 宿主: 兔 Polyclonal RB37199 unconjugated
产品编号 ABIN1537116
发货至: 中国
  • 抗原 See all MR1 抗体
    MR1 (Major Histocompatibility Complex, Class I-Related (MR1))
    抗原表位
    • 10
    • 7
    • 6
    • 4
    • 2
    • 2
    • 1
    • 1
    • 1
    AA 312-341, C-Term
    适用
    • 34
    • 2
    • 1
    • 1
    宿主
    • 30
    • 5
    克隆类型
    • 31
    • 4
    多克隆
    标记
    • 15
    • 4
    • 3
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This MR1 antibody is un-conjugated
    应用范围
    • 24
    • 23
    • 5
    • 3
    • 3
    • 1
    • 1
    • 1
    Western Blotting (WB)
    纯化方法
    This antibody is purified through a protein A column, followed by peptide affinity purification.
    免疫原
    This MR1 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 312-341 amino acids from the C-terminal region of human MR1.
    克隆位点
    RB37199
    亚型
    IgG
    Top Product
    Discover our top product MR1 Primary Antibody
  • 应用备注
    WB: 1:1000
    限制
    仅限研究用
  • 生效 #101753 (Immunocytochemistry)
    '独立验证'标志
    by
    Dr. Randy Brutkiewicz Laboratory, Department of Microbiology and Immunology, Indiana University School of Medicine
    No.
    #101753
    日期
    2018.02.20
    抗原
    MR1
    Lot Number
    SA111213CH
    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 ABIN1537116 specifically labels the targeted antigen in HEK293 ectopically expressing human MR1 in ICC.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1537116
    Secondary Antibody
    Texas Red-conjugated donkey anti-rabbit immunoglobulin antiserum (Jackson ImmunoResearch, 711-076-152, lot 66576)
    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 cells in sterile glass-bottom 35-mm dishes coated with collagen (MatTek, P35GCol-1.5-14-C). Let cells grow to 50-80% confluency.
    • Wash cells d with 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 atRT.
    • Incubate cells with primary rabbit anti-MR1 antibody (antibodies-online, ABIN1537116, lot SA111213CH) 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.
    • Incubate cells with Texas Red-conjugated donkey anti-rabbit immunoglobulin antiserum (Jackson ImmunoResearch, 711-076-152, lot 66576) diluted 1:50 in dilution buffer for 1h at RT.
    • Wash cells 3x with PBS.
    • To stain the nucleus, cells were immersed in PBS-containing Hoechst diluted 1:2000 in PBS 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 ABIN1537116 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 #102828 (Immunoprecipitation)
    '独立验证'标志
    by
    Dr. Randy Brutkiewicz Laboratory, Department of Microbiology and Immunology, Indiana University School of Medicine
    No.
    #102828
    日期
    2018.02.20
    抗原
    MR1
    Lot Number
    SA111213CH
    Method validated
    Immunoprecipitation
    Positive Control
    HEK293 cells transfected with human MR1 cDNA
    Negative Control
    HEK293 cells transfected with plasmid vector only
    Notes

    Passed. ABIN1537116 immunoprecipitates human MR1 overexpressed by HEK293 cells.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1537116
    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 ABIN1537116, but not the isotype control, immunoprecipitates with human MR1 overexpressed by HEK293 cells.

  • 生效 #102829 (Western Blotting)
    '独立验证'标志
    by
    Dr. Randy Brutkiewicz Laboratory, Department of Microbiology and Immunology, Indiana University School of Medicine
    No.
    #102829
    日期
    2018.02.20
    抗原
    MR1
    Lot Number
    SA111213CH
    Method validated
    Western Blotting
    Positive Control
    HEK293 cells transfected with human MR1 cDNA
    Negative Control
    HEK293 cells transfected with plasmid vector only
    Notes

    Passed. ABIN1537116 recognizes human MR1 overexpressed by HEK293 cells in a western blot.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1537116
    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).
    • Denature 200µg total protein for 5min at 95°C in 20µl 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.
      • loading control rabbit anti beta-actin (antibodies-online, ABIN962807) diluted 1:500 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 10min with PBS/0.05% Tween-20.
    • Reveal protein bands using an Odyssey imaging system (LI-COR Biosciences).
    Experimental Notes

    The human MR1 antibody ABIN1537116 reveals a protein of the expected molecular weight of MR1 in lysates of human MR1-expressing HEK293 cells. The protein bands is only visible in the positive but not the negative controls.

  • 状态
    Liquid
    缓冲液
    Purified polyclonal antibody supplied in PBS with 0.09 % (W/V) sodium azide.
    储存液
    Sodium azide
    注意事项
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    储存条件
    4 °C,-20 °C
    储存方法
    MR1 Antibody (C-term) can be refrigerated at 2-8 °C for up to 6 months. For long term storage, keep at -20 °C.
    有效期
    6 months
  • 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
    MR1 has antigen presentation function. Involved in the development and expansion of a small population of T cells expressing an invariant T cell receptor alpha chain called mucosal-associated invariant T cells (MAIT). MAIT cells are preferentially located in the gut lamina propria and therfore may be involed in monitoring commensal flora or serve as a distress signal. Expression and MAIT cell recognition seem to be ligand-dependent.
    分子量
    39366
    基因ID
    3140
    NCBI登录号
    NP_001181928, NP_001181929, NP_001181964, NP_001522
    UniProt
    Q95460
    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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