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Signal Transduction Signaling Pathway Nuclear Signaling STATs

人STAT3 knockout HeLa cell line (ab255436)

  • Datasheet
  • SDS
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Western blot - Human STAT3 knockout HeLa cell line (ab255436)
  • Sanger Sequencing - Human STAT3 knockout HeLa cell line (ab255436)

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概述

  • 产品名称

    人STAT3 knockout HeLa cell line
    参阅全部 STAT3 细胞裂解液
  • Parental Cell Line

    HeLa
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2
  • Passage number

    <20
  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • 经测试应用

    适用于: WBmore details
  • Biosafety level

    2
  • 常规说明

    Recommended control: Human wild-type HeLa cell line (ab255448). 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
  • Adherent /Suspension

    Adherent
  • Tissue

    Cervix
  • Cell type

    epithelial
  • Disease

    Adenocarcinoma
  • Gender

    Female
  • STR Analysis

    Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10
  • 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

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • STATs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Stem Cells
    • Embryonic Stem Cells
    • Intracellular
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • STATs
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • STAT family
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Transcription factors
    • Developmental Biology
    • Embryogenesis
    • Embryonic stem cells
    • Surface molecules

靶标

  • 功能

    Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene (PubMed:17344214). May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1 (PubMed:17344214). Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation (PubMed:18242580). Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity.
  • 组织特异性

    Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
  • 疾病相关

    Hyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
    Autoimmune disease, multisystem, infantile-onset
  • 序列相似性

    Belongs to the transcription factor STAT family.
    Contains 1 SH2 domain.
  • 翻译后修饰

    Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling.
  • 细胞定位

    Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3. Identified in a complex with LYN and PAG1.
  • Target information above from: UniProt accession P40763 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 STAT3 knockout HeLa cell lysate (ab263797)
  • Related Products

    • Anti-STAT3 antibody [EPR361] (ab109085)
    • Anti-STAT3 antibody [EPR787Y] - BSA and Azide free (ab171359)
    • Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    • Anti-STAT3 (phospho Y705) antibody [EPR23968-52] (ab267373)
    • Anti-STAT3 (phospho Y705) antibody [EPR23968-52] - BSA and Azide free (ab280202)
    • Anti-STAT3 antibody [EPR787Y] (ab68153)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab255436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: 88 kDa.
说明
WB
Use at an assay dependent concentration. Predicted molecular weight: 88 kDa.

图片

  • Western blot - Human STAT3 knockout HeLa cell line (ab255436)
    Western blot - Human STAT3 knockout HeLa cell line (ab255436)
    All lanes : Anti-STAT3 (phospho Y705) antibody [EPR23968-52] (ab267373) at 1/1000 dilution

    Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) serum starved overnight, then treated with 50 ng/ml IFN alpha for 30 minutes, whole cell lysate
    Lane 2 : STAT3 knockout HeLa serum starved overnight, then treated with 50 ng/ml IFN alpha for 30 minutes, whole cell lysate
    Lane 3 : Jurkat (human t cell leukemia cell line from peripheral blood) treated with 50 ng/ml IFN alpha for 30 minutes, whole cell lysate
    Lane 4 : HepG2 (human hepatocellar carcinoma epithelial cell) serum starved overnight, then treated with 100 ng/ml IL-6 for 30 minutes, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution

    Predicted band size: 88 kDa
    Observed band size: 88 kDa



    Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

    Lanes 1-4: Merged signal (red and green). Green - ab267373 observed at 88 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

    Lanes 1-2: ab267373 Anti-STAT3 (phospho Y705) antibody [EPR23968-52] was shown to specifically react with STAT3 in wild-type serum starved and then IFN alpha treated HeLa cells. Loss of signal was observed when serum starved and then IFN alpha treated knockout cell line ab255436 (knockout cell lysate ab263797) was used. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. ab267373 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

    Lysates loaded onto lanes 3-4 were made freshly and used in WB immediately to minimize protein degradation.

  • Sanger Sequencing - Human STAT3 knockout HeLa cell line (ab255436)
    Sanger Sequencing - Human STAT3 knockout HeLa cell line (ab255436)
    Homozygous: 1 bp 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)

发表研究结果有使用 ab255436?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab255436 尚未被引用在任何文献中。

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