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Cardiovascular Angiogenesis Cytokines Interleukin

人CXCL8 knockout PC-3 cell line (ab273743)

  • Datasheet
  • SDS
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Western blot - Human CXCL8 knockout PC3 cell line (ab273743)
  • Sandwich ELISA - Human CXCL8 knockout PC-3 cell line (ab273743)
  • Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)
  • Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)
  • Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)

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概述

  • 产品名称

    人CXCL8 knockout PC-3 cell line
  • Parental Cell Line

    PC3
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 22% 11 bp deletion, 24% 7 bp deletion, 54% 2 bp deletion in exon 2
  • Passage number

    <20
  • Knockout validation

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

    适用于: WB, Sandwich ELISAmore details
  • Biosafety level

    1
  • 常规说明

    Recommended control: Human wild-type PC3 cell line (ab275472). 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: F-12K + 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
  • Adherent /Suspension

    Adherent
  • Tissue

    Prostate
  • Cell type

    epithelial
  • Disease

    Adenocarcinoma
  • 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

    • Cardiovascular
    • Angiogenesis
    • Cytokines
    • Interleukin
    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Neuroscience
    • Processes

靶标

  • 功能

    IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T-cells, but not monocytes. It is also involved in neutrophil activation. It is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively.
  • 序列相似性

    Belongs to the intercrine alpha (chemokine CxC) family.
  • 翻译后修饰

    Several N-terminal processed forms are produced by proteolytic cleavage after secretion from at least peripheral blood monocytes, leukcocytes and endothelial cells. In general, IL-8(1-77) is referred to as interleukin-8. IL-8(6-77) is the most promiment form.
  • 细胞定位

    Secreted.
  • Target information above from: UniProt accession P10145 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt

相关产品

  • Biochemicals

    • Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • KO cell lysates

    • Human CXCL8 knockout PC-3 cell lysate (ab275520)
  • Related Products

    • Anti-IL-8 antibody [EPR22994-255] (ab235584)
    • Anti-IL-8 antibody [EPR22994-255] - BSA and Azide free (ab275255)
  • SimpleStep ELISA kits

    • Human IL-8 ELISA Kit (ab214030)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab273743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.
Sandwich ELISA
Use at an assay dependent concentration.
说明
WB
Use at an assay dependent concentration.
Sandwich ELISA
Use at an assay dependent concentration.

图片

  • Western blot - Human CXCL8 knockout PC3 cell line (ab273743)
    Western blot - Human CXCL8 knockout PC3 cell line (ab273743)
    All lanes : Anti-IL-8 antibody [EPR22994-255] (ab235584) at 1/1000 dilution

    Lane 1 : Wild-type PC-3 Brefeldin A (ab120299)-treated (5 μg/ml, 5 h) cell lysate
    Lane 2 : Wild-type PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (ab120299) (5 μg/ml, 5 h) cell lysate
    Lane 3 : CXCL8 knockout PC-3 Brefeldin A (ab120299)-treated (5 μg/ml, 5 h) cell lysate
    Lane 4 : CXCL8 knockout PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (ab120299) (5 μg/ml, 5 h) cell lysate
    Lane 5 : A431 cell lysate
    Lane 6 : HCT116 cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Observed band size: 10 kDa why is the actual band size different from the predicted?



    Lanes 1 - 6: Merged signal (red and green). Green - ab235584 observed at 10 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab235584 was shown to react with IL-8 in wild-type PC-3 cells in Western blot with loss of signal observed in CXCL8 knockout cell line ab273743 (knockout cell lysate ab275520). Wild-type PC-3 and CXCL8 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab235584 and ab8245 (Mouse anti-GAPDH antibody [6C5]) 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.

  • Sandwich ELISA - Human CXCL8 knockout PC-3 cell line (ab273743)
    Sandwich ELISA - Human CXCL8 knockout PC-3 cell line (ab273743)

    Human IL-8 concentration was interpolated from the standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human IL-8 ELISA kit (ab214030). Wild-type PC-3 cells and CXCL8 knockout PC-3 cells (ab273743) were assessed in duplicate (n=2) and were either treated with 2 µg/ml LPS for 6 hours to induce expression of IL-8 or not treated with LPS. Data are represented as the mean and error bars represent standard deviation.

  • Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)
    Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)

    Allele-1: 11bp deletion in exon 2.

  • Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)
    Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)

    Allele-2: 7bp deletion in exon 2.

  • Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)
    Next Generation Sequencing - Human CXCL8 (IL-8) knockout PC3 cell line (ab273743)

    Allele-3: 2bp deletion in exon 2.

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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