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    products/cell-lines/human-sox17-knockout-hela-cell-line-ab265744.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families HMG Box
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Human SOX17 knockout HeLa cell line (ab265744)

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Immunocytochemistry/ Immunofluorescence - Human SOX17 knockout HeLa cell line (ab265744)
  • Western blot - Human SOX17 knockout HeLa cell line (ab265744)
  • Sanger Sequencing - Human SOX17 knockout HeLa cell line (ab265744)
  • Sanger Sequencing - Human SOX17 knockout HeLa cell line (ab265744)

You may also be interested in

Primary
Product image
Anti-SOX17 antibody [EPR20684] (ab224637)

View more associated products

Overview

  • Product name

    Human SOX17 knockout HeLa cell line
    See all SOX17 lysates
  • Parental Cell Line

    HeLa
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 8 bp deletion in exon 1
  • Passage number

    <20
  • Knockout validation

    Immunocytochemistry (ICC), Sanger Sequencing, Western Blot (WB)
  • Tested applications

    Suitable for: ICC, WBmore details
  • Biosafety level

    2
  • General notes

    Recommended control: Human wild-type HeLa cell line (ab255928). 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.

Properties

  • Number of cells

    1 x 106 cells/vial, 1 mL
  • Viability

    ~80%
  • 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
  • Antibiotic resistance

    Puromycin 1.00µg/ml
  • Mycoplasma free

    Yes
  • Storage instructions

    Shipped on Dry Ice. Store in liquid nitrogen.
  • Storage buffer

    Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HMG Box
    • Stem Cells
    • Embryonic Stem Cells
    • Intracellular
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Developmental Biology
    • Embryogenesis
    • Embryonic stem cells
    • Surface molecules

Target

  • Function

    Acts as transcription regulator that binds target promoter DNA and bends the DNA. Binds to the sequences 5'-AACAAT-'3 or 5'-AACAAAG-3'. Modulates transcriptional regulation via WNT3A. Inhibits Wnt signaling. Promotes degradation of activated CTNNB1. Plays a key role in the regulation of embryonic development. Required for normal looping of the embryonic heart tube. Required for normal development of the definitive gut endoderm. Probable transcriptional activator in the premeiotic germ cells.
  • Tissue specificity

    Expressed in adult heart, lung, spleen, testis, ovary, placenta, fetal lung, and kidney. In normal gastrointestinal tract, it is preferentially expressed in esophagus, stomach and small intestine than in colon and rectum.
  • Involvement in disease

    Defects in SOX17 are the cause of vesicoureteral reflux type 3 (VUR3) [MIM:613674]. VUR3 is a disease belonging to the group of congenital anomalies of the kidney and urinary tract. It is characterized by the reflux of urine from the bladder into the ureters and sometimes into the kidneys, and is a risk factor for urinary tract infections. Primary disease results from a developmental defect of the ureterovesical junction. In combination with intrarenal reflux, the resulting inflammatory reaction may result in renal injury or scarring, also called reflux nephropathy. Extensive renal scarring impairs renal function and may predispose patients to hypertension, proteinuria, renal insufficiency and end-stage renal disease.
  • Sequence similarities

    Contains 1 HMG box DNA-binding domain.
    Contains 1 Sox C-terminal domain.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession Q9H6I2 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt

Associated products

  • KO cell lysates

    • Human SOX17 knockout HeLa cell lysate (ab257697)
  • Related Products

    • Anti-SOX17 antibody [EPR20684] (ab224637)
    • Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab265744 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 44 kDa.
Notes
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 44 kDa.

Images

  • Immunocytochemistry/ Immunofluorescence - Human SOX17 knockout HeLa cell line (ab265744)
    Immunocytochemistry/ Immunofluorescence - Human SOX17 knockout HeLa cell line (ab265744)
    ab224637 staining SOX17 in wild-type HeLa cells (top panel) and SOX17 knockout HeLa cells (ab265744) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 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 ab224637 at 0.2μg/ml concentration 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).
  • Western blot - Human SOX17 knockout HeLa cell line (ab265744)
    Western blot - Human SOX17 knockout HeLa cell line (ab265744)
    All lanes : Anti-SOX17 antibody [EPR20684] (ab224637) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SOX17 knockout HeLa cell lysate
    Lane 3 : SK-OV-3 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 44 kDa
    Observed band size: 51 kDa why is the actual band size different from the predicted?



    Lanes 1-3: Merged signal (red and green). Green - ab224637 observed at 51 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab224637 Anti-SOX17 antibody [EPR20684] was shown to specifically react with SOX17 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265744 (knockout cell lysate ab257697) was used.  Wild-type and SOX17 knockout samples were subjected to SDS-PAGE.  ab224637 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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 20000 dilution for 1 hour at room temperature before imaging.

  • Sanger Sequencing - Human SOX17 knockout HeLa cell line (ab265744)
    Sanger Sequencing - Human SOX17 knockout HeLa cell line (ab265744)

    Allele-1: 8 bp deletion in exon 1.

     

  • Sanger Sequencing - Human SOX17 knockout HeLa cell line (ab265744)
    Sanger Sequencing - Human SOX17 knockout HeLa cell line (ab265744)

    Allele-2: 2 bp deletion in exon 1.

     

Protocols

  • Hemocytometer protocol
  • Mammalian cell tissue culture techniques protocol

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab265744? Please let us know so that we can cite the reference in this datasheet.

ab265744 has not yet been referenced specifically in any publications.

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