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Validated using a knockout cell lineRecombinantRabMAb

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

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Immunocytochemistry/ Immunofluorescence - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
  • Western blot - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
  • Immunoprecipitation - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
  • Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20684] to SOX17 - BSA and Azide free
  • Suitable for: ICC/IF, IHC-P, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-SOX17 antibody [EPR20684] - BSA and Azide free
    See all SOX17 primary antibodies

  • Description

    Rabbit monoclonal [EPR20684] to SOX17 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IHC-P, IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Human seminoma tissue. IP: SK-OV-3 cell lysate. WB: HeLa and SK-OV-3 cell lysates. ICC/IF: HeLa cells.

  • General notes

    ab226862 is the carrier-free version of ab224637.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab226862 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/IF
Use a concentration of 0.2 µg/ml.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 44 kDa.
Notes
ICC/IF
Use a concentration of 0.2 µg/ml.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 44 kDa.

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.
  • Information by UniProt

  • Database links

    • Alternative names

      • FLJ22252 antibody
      • SOX17 antibody
      • SOX17_HUMAN antibody
      • SRY (sex determining region Y) box 17 antibody
      • SRY box 17 antibody
      • SRY related HMG box transcription factor SOX17 antibody
      • Transcription factor SOX-17 antibody
      • Transcription factor SOX17 antibody
      • VUR3 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
      Immunocytochemistry/ Immunofluorescence - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
      This data was developed using the same antibody clone in a different buffer formulation (ab224637). 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 - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
      Western blot - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
      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?



      This data was developed using the same antibody clone in a different buffer formulation (ab224637).

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)

      Immunohistochemical analysis of paraffin-embedded human seminoma tissue labeling SOX17 with ab224637 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on tumor cells of human seminoma (PMID:19369635; PMID:18348160) is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224637).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

      SOX17 was immunoprecipitated from 0.35 mg of SK-OV-3 (human ovarian cancer cell line) lysate with ab224637 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224637 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Lane 1: SK-OV-3 whole cell lysate 10 µg (Input). 

      Lane 2: ab224637 IP in SK-OV-3 whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab224637 in SK-OV-3 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 1 second.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224637).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)

      Immunohistochemical analysis of paraffin-embedded human choriocarcinoma tissue labeling SOX17 with ab224637 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Negative tissue: no staining on tumor cells of human choriocarcinoma (PMID: 19369635) is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224637).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)

      Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling SOX17 with ab224637 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on endothelium of rat lung (PMID:24418654) is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224637).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] - BSA and Azide free (ab226862)

      Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling SOX17 with ab224637 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on endothelium of mouse spleen (PMID:24418654) is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224637).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    Protocols

    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

    References (0)

    Publishing research using ab226862? Please let us know so that we can cite the reference in this datasheet.

    ab226862 has not yet been referenced specifically in any publications.

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