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Immunology Adaptive Immunity B Cells CD

人CD80 knockout Raji cell line (ab273873)

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  • SDS
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Next Generation Sequencing - Human CD80 knockout Raji cell line (ab273873)
  • Western blot - Human CD80 knockout Raji cell line (ab273873)
  • Next Generation Sequencing - Human CD80 knockout Raji cell line (ab273873)

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概述

  • 产品名称

    人CD80 knockout Raji cell line
    参阅全部 CD80 细胞裂解液
  • Parental Cell Line

    Raji
  • Organism

    Human
  • Mutation description

    Knockout achieved by CRISPR/Cas9; X = 5 bp deletion; Frameshift: 100%
  • Passage number

    <20
  • Knockout validation

    Next Generation Sequencing (NGS), Western Blot (WB)
  • 经测试应用

    适用于: Next Generation Sequencing, WBmore details
  • Biosafety level

    2
  • 常规说明

    Recommended control: Human wild-type Raji cell line (ab271145). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

    Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

    Culture medium: RPMI + 10% FBS

    Initial handling guidelines: Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

    1. Thaw the vial in 37°C water for bath approximately 1-2 minutes.
    2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
    3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
    4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.

    Initial handling guidelines: 

    • All seeding densities should be based on cell counts gained by established methods.
    • A guide seeding density of 4x105 cells/mL is recommended.
    • A maximum of 3x106 viable cells/ml is obtainable.

    This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

    We will provide viable cells that proliferate on revival.

性能

  • Number of cells

    1 x 106 cells/vial, 1 mL
  • Adherent /Suspension

    Suspension
  • Tissue

    Lymphatic
  • Cell type

    Burkitt's lymphoma
  • Disease

    Lymphoma
  • Gender

    Male
  • Mycoplasma free

    Yes
  • 存放说明

    Shipped on Dry Ice. Store in liquid nitrogen.
  • 存储溶液

    Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether
  • 研究领域

    • Immunology
    • Adaptive Immunity
    • B Cells
    • CD
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Dendritic Cell Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Monocytic Lineage

靶标

  • 功能

    Involved in the costimulatory signal essential for T-lymphocyte activation. T-cell proliferation and cytokine production is induced by the binding of CD28 or CTLA-4 to this receptor.
  • 组织特异性

    Expressed on activated B-cells, macrophages and dendritic cells.
  • 序列相似性

    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • 细胞定位

    Membrane.
  • Target information above from: UniProt accession P33681 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt

相关产品

  • KO cell lysates

    • Human CD80 knockout Raji cell lysate (ab273827)
  • Related Products

    • Anti-CD80 antibody [EPR1157(2)] (ab134120)
    • Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab273873于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Next Generation Sequencing
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
说明
Next Generation Sequencing
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.

图片

  • Next Generation Sequencing - Human CD80 knockout Raji cell line (ab273873)
    Next Generation Sequencing - Human CD80 knockout Raji cell line (ab273873)
    5 bp deletion after Glu85 of the WT protein
  • Western blot - Human CD80 knockout Raji cell line (ab273873)
    Western blot - Human CD80 knockout Raji cell line (ab273873)
    All lanes : Anti-CD80 antibody [EPR1157(2)] (ab134120) at 1/1000 dilution

    Lane 1 : Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 2 : CD80 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 3 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.


    Lanes 1 - 3: Merged signal (red and green). Green - ab134120 observed at 55 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

     ab134120 was shown to react with CD80 in wild-type Raji cells in western blot with loss of signal observed in CD80 knockout cell line ab273873 (knockout cell lysate ab273827). Wild-type and CD80 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134120 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Next Generation Sequencing - Human CD80 knockout Raji cell line (ab273873)
    Next Generation Sequencing - Human CD80 knockout Raji cell line (ab273873)

    Knockout achieved by CRISPR/Cas9; X = 5 bp deletion; Frameshift: 100%

     

     

实验方案

  • Hemocytometer protocol
  • Mammalian cell tissue culture techniques protocol

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (0)

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