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Immunology Innate Immunity Cytokines Interleukins

人IL13RA2 knockout A375 cell line (ab273381)

  • Datasheet
  • SDS
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Western blot - Human IL13RA2 knockout A375 cell line (ab273381)
  • Immunocytochemistry/ Immunofluorescence - Human IL13RA2 knockout A375 cell line (ab273381)
  • Next Generation Sequencing - Human IL13RA2 knockout A375 cell line (ab273381)

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概述

  • 产品名称

    人IL13RA2 knockout A375 cell line
    参阅全部 IL-13 receptor alpha 2 细胞裂解液
  • Parental Cell Line

    A375
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3
  • Passage number

    <20
  • Knockout validation

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

    适用于: ICC, WBmore details
  • Biosafety level

    1
  • 常规说明

    Recommended control: Human wild-type A375 cell line (ab275461). 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: DMEM (High Glucose) + 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 bath for 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 2x104 cells/cm2. 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.

    Subculture guidelines:

    • All seeding densities should be based on cell counts gained by established methods.
    • A guide seeding density of 2x104 cells/cm2 is recommended.
    • A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
    • Cells should be passaged when they have achieved 80-90% confluence.

    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
  • Viability

    ~80%
  • Adherent /Suspension

    Adherent
  • Tissue

    Skin
  • Cell type

    epithelial
  • Disease

    Melanoma
  • Gender

    Female
  • 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
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Immunology
    • Innate Immunity
    • Cytokines
    • Other
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus

靶标

  • 功能

    Binds as a monomer with high affinity to interleukin-13 (IL13), but not to interleukin-4 (IL4).
  • 序列相似性

    Belongs to the type I cytokine receptor family. Type 5 subfamily.
    Contains 1 fibronectin type-III domain.
  • 结构域

    The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
    The box 1 motif is required for JAK interaction and/or activation.
  • 细胞定位

    Membrane.
  • Target information above from: UniProt accession Q14627 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 IL13RA2 knockout A375 cell lysate (ab275532)
  • Related Products

    • Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044)
    • Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] - BSA and Azide free (ab263874)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab273381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
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
说明
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.

图片

  • Western blot - Human IL13RA2 knockout A375 cell line (ab273381)
    Western blot - Human IL13RA2 knockout A375 cell line (ab273381)
    All lanes : Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044) at 1/1000 dilution

    Lane 1 : Wild-type A375 cell lysate
    Lane 2 : IL13RA2 knockout A375 cell lysate
    Lane 3 : U-87-MG cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Observed band size: 49 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab260044 observed at 49 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab260044 was shown to react with IL-13 receptor alpha 2 in wild-type A375 cells in western blot with loss of signal observed in IL13RA2 knockout cell line ab273371 (knockout cell lysate ab275532). Wild-type and IL13RA2 knockout A375 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab260044 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.

  • Immunocytochemistry/ Immunofluorescence - Human IL13RA2 knockout A375 cell line (ab273381)
    Immunocytochemistry/ Immunofluorescence - Human IL13RA2 knockout A375 cell line (ab273381)
    ab260044 staining IL-13 receptor alpha 2 in wild-type A375 cells (top panel) and IL13RA2 knockout A375 cells (bottom panel) (ab273381). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab260044 at 1/500 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
  • Next Generation Sequencing - Human IL13RA2 knockout A375 cell line (ab273381)
    Next Generation Sequencing - Human IL13RA2 knockout A375 cell line (ab273381)

    Homozygous: 1 bp deletion in exon 3

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)

发表研究结果有使用 ab273381?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab273381 尚未被引用在任何文献中。

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