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人CD74 knockout Raji cell裂解物(ab273830)

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Next Generation Sequencing - Human CD74 knockout Raji cell lysate (ab273830)
  • Western blot - Human CD74 knockout Raji cell lysate
  • Western blot - Human CD74 knockout Raji cell lysate

概述

  • 产品名称

    人CD74 knockout Raji cell裂解物
    参阅全部 CD74 试剂盒

  • 产品概述

     Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    Raji

  • Organism

    Human

  • Mutation description

    Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift: 100%

  • Passage number

    <20

  • Knockout validation

    Next Generation Sequencing (NGS), Western Blot (WB)

  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • 说明

    Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.

    User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

    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.

  • 经测试应用

    适用于: WBmore details

性能

靶标

  • 功能

    Plays a critical role in MHC class II antigen processing by stabilizing peptide-free class II alpha/beta heterodimers in a complex soon after their synthesis and directing transport of the complex from the endoplasmic reticulum to the endosomal/lysosomal system where the antigen processing and binding of antigenic peptides to MHC class II takes place. Serves as cell surface receptor for the cytokine MIF.

  • 序列相似性

    Contains 1 thyroglobulin type-1 domain.

  • 细胞定位

    Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network. Endosome. Lysosome. Transits through a number of intracellular compartments in the endocytic pathway. It can either undergo proteolysis or reach the cell membrane.
  • Information by UniProt

  • 别名

    • CD 74
    • CD74
    • CD74 antigen
    • CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated)
    • CD74 molecule
    • CD74 molecule, major histocompatibility complex, class II invariant chain
    • CLIP
    • DHLAG
    • Gamma chain of class II antigens
    • HG2A_HUMAN
    • HLA class II histocompatibility antigen gamma chain
    • HLA DR antigens associated invariant chain
    • HLA DR gamma
    • HLA-DR antigens-associated invariant chain
    • HLA-DR-gamma
    • HLADG
    • HLADR antigens associated invariant chain
    • Ia antigen associated invariant chain
    • Ia antigen-associated invariant chain
    • Ia associated invariant chain
    • Ia gamma
    • Ii
    • Invariant polypeptide of major histocompatibility complex class II antigen associated
    • la-gamma
    • Major histocompatibility complex class II invariant chain
    • MHC HLA DR gamma chain
    • MHC HLA-DR gamma chain
    • p33
    • p35
    • Protein 41
    see all

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab273830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
WB
Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.
说明
WB
Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.

图片

  • Next Generation Sequencing - Human CD74 knockout Raji cell lysate (ab273830)
    Next Generation Sequencing - Human CD74 knockout Raji cell lysate (ab273830)
    Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift: 100%
  • Western blot - Human CD74 knockout Raji cell lysate
    Western blot - Human CD74 knockout Raji cell lysate

    Lane 1: Raji (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
    Lane 2:     CD74 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
    Lane 3:     Daudi (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
    Lane 4:     Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)

    Lanes 1 - 4: Merged signal (red and green). Green - ab108393 observed at 34 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab108393 was shown to react with CD74 in wild-type Raji cells in Western blot with loss of signal observed in CD74 knockout cell line ab273876 (knockout cell lysate ab273830). Wild-type Raji and CD74 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab108393 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 h at room temperature before imaging.

  • Western blot - Human CD74 knockout Raji cell lysate
    Western blot - Human CD74 knockout Raji cell lysate

    Lane 1: Raji (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
    Lane 2:     CD74 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
    Lane 3:     Daudi (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
    Lane 4:     Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)

    Lanes 1 - 4: Merged signal (red and green). Green - ab64772 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab64772 was shown to react with CD74 in wild-type Raji cells in Western blot with loss of signal observed in CD74 knockout cell line ab273876 (knockout cell lysate ab273830). Wild-type Raji and CD74 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab64772 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 µg/ml 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 h at room temperature before imaging.

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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