Recombinant
RabMAb

Recombinant Anti-SCF antibody [EP665Y] (ab52603)

Reviews (5)Q&A (2)References (10)

Overview

  • Product name

    Anti-SCF antibody [EP665Y]
    See all SCF primary antibodies

  • Description

    Rabbit monoclonal [EP665Y] to SCF

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human SCF aa 50-150 (N terminal). The exact sequence is proprietary.
    Database link: P21583

  • Positive control

    • Human kidney carcinoma tissue.

  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52603 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
WB 1/10000. Predicted molecular weight: 31 kDa.
Flow Cyt 1/20 - 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/500.

Target

  • Function

    Ligand for the receptor-type protein-tyrosine kinase KIT. Plays an essential role in the regulation of cell survival and proliferation, hematopoiesis, stem cell maintenance, gametogenesis, mast cell development, migration and function, and in melanogenesis. KITLG/SCF binding can activate several signaling pathways. Promotes phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase, and subsequent activation of the kinase AKT1. KITLG/SCF and KIT also transmit signals via GRB2 and activation of RAS, RAF1 and the MAP kinases MAPK1/ERK2 and/or MAPK3/ERK1. KITLG/SCF and KIT promote activation of STAT family members STAT1, STAT3 and STAT5. KITLG/SCF and KIT promote activation of PLCG1, leading to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate. KITLG/SCF acts synergistically with other cytokines, probably interleukins.

  • Involvement in disease

    Hyperpigmentation with or without hypopigmentation, familial progressive
    Deafness, congenital, unilateral or asymmetric

  • Sequence similarities

    Belongs to the SCF family.

  • Developmental stage

    Acts in the early stages of hematopoiesis.

  • Post-translational
    modifications

    A soluble form (sKITLG) is produced by proteolytic processing of isoform 1 in the extracellular domain.
    Found in two differentially glycosylated forms, LMW-SCF and HMW-SCF. LMW-SCF is fully N-glycosylated at Asn-145, partially N-glycosylated at Asn-90, O-glycosylated at Ser-167, Thr-168 and Thr-180, and not glycosylated at Asn-97 or Asn-118. HMW-SCF is N-glycosylated at Asn-118, Asn-90 and Asn-145, O-glycosylated at Ser-167, Thr-168 and Thr-180, and not glycosylated at Asn-97.
    A soluble form exists as a cleavage product of the extracellular domain.

  • Cellular localization

    Secreted; Cell membrane and Cytoplasm. Cytoplasm, cytoskeleton. Cell membrane. Cell projection, lamellipodium. Cell projection, filopodium.
  • Information by UniProt

  • Database links

    • Alternative names

      • C kit ligand antibody
      • C-kit ligand antibody
      • Ckit ligand antibody
      • DCUA antibody
      • DFNA69 antibody
      • DKFZp686F2250 antibody
      • familial progressive hyperpigmentation 2 antibody
      • FPH2 antibody
      • FPHH antibody
      • KIT ligand antibody
      • Kitl antibody
      • KITLG antibody
      • KL 1 antibody
      • KL1 antibody
      • Mast cell growth factor antibody
      • MGF antibody
      • MGF stem cell factor antibody
      • SCF antibody
      • SCF_HUMAN antibody
      • SF antibody
      • SHEP7 antibody
      • sKITLG antibody
      • Soluble KIT ligand antibody
      • Steel factor antibody
      • steel, mouse, homolog of antibody
      • Stem cell factor antibody
      • Stem cell factor precursor antibody
      see all

    Images

    • Western blot - Anti-SCF antibody [EP665Y] (ab52603)
      Western blot - Anti-SCF antibody [EP665Y] (ab52603)
      Anti-SCF antibody [EP665Y] (ab52603) at 1/10000 dilution + recombinant protein (5ng)

      Secondary
      HRP-labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 31 kDa
      Observed band size: 18 kDa
      why is the actual band size different from the predicted?

    • Immunocytochemistry/ Immunofluorescence - Anti-SCF antibody [EP665Y] (ab52603)
      Immunocytochemistry/ Immunofluorescence - Anti-SCF antibody [EP665Y] (ab52603)

      Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling SCF with purified ab52603 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCF antibody [EP665Y] (ab52603)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCF antibody [EP665Y] (ab52603)

      Immunohistochemical analysis of paraffin-embedded human kidney carcinoma tissue using ab52603 at a dilution of 1/100-1/250.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Flow Cytometry - Anti-SCF antibody [EP665Y] (ab52603)
      Flow Cytometry - Anti-SCF antibody [EP665Y] (ab52603)
      Overlay histogram showing MCF-7 cells stained with ab52603 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52603, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min) used under the same conditions.

      Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

    References

    This product has been referenced in:

    • Frumento G  et al. CD117 (c-Kit) Is Expressed During CD8+ T Cell Priming and Stratifies Sensitivity to Apoptosis According to Strength of TCR Engagement. Front Immunol 10:468 (2019). Read more (PubMed: 30930902) »
    • Yong KS  et al. Human CD34(lo)CD133(lo) fetal liver cells support the expansion of human CD34(hi)CD133(hi) hematopoietic stem cells. Cell Mol Immunol 13:605-14 (2016). ICC/IF . Read more (PubMed: 27593483) »
    See all 10 Publications for this product

    Publishing research using ab52603? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-7 of 7 Abreviews or Q&A

    Western blot abreview for Anti-SCF antibody [EP665Y]

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (Canine cell lines, tissues and recombinant canine)
    Gel Running Conditions
    Reduced Denaturing (7.5%)
    Loading amount
    50 µg
    Specification
    Canine cell lines, tissues and recombinant canine
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 16 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-SCF antibody [EP665Y]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (NIH3T3)
    Permeabilization
    Yes - 1%tween
    Specification
    NIH3T3
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 25 2015

    Western blot abreview for Anti-SCF antibody [EP665Y]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    5 µg
    Gel Running Conditions
    Non-reduced Denaturing (14%)
    Sample
    Human Cell lysate - whole cell (CLL cells)
    Specification
    CLL cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 4% · Temperature: 25°C
    Read More

    Mr. George Gavriilidis

    Verified customer

    Submitted Feb 26 2015

    Western blot abreview for Anti-SCF antibody [EP665Y]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (12%)
    Sample
    Human Serum (serum)
    Specification
    serum
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 31 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-SCF antibody [EP665Y]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 10 minute(s) · Concentration: 5%
    Sample
    Human Cell (H9 embryonic stem cells)
    Specification
    H9 embryonic stem cells
    Permeabilization
    Yes - 0.01% Triton X100
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 22 2014

    Question



    Inquiry: Does this antibody recognize both the membrane bound and soluble SCF?

    Read More
    Answer

    Thank you for contacting us.


    Based on the immunogen sequence, the antibody could recognize the soluble SCF and the extracellular domain.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    -----------------------------------------------------------------------------------------------

    Do you wish running an ELISA was fast and easy? Check out our new SimpleStep™ ELISA, a single wash, colorimetric sandwich ELISA assay.

    Discover more at www.abcam.com/simplestep

    Read More

    Question

    I have a couple of questions regarding this antibody: 1. What was the exact sequence from SCF that was used as the immunogen? 2. Has this antibody been used for flow cytometry and have the results been published? Thanks!

    Read More
    Answer

    Thank you for your enquiry. Unfortunately, the immunogen sequence for this product is proprietary information. I can confirm that this product works in flow cytometry. However, we currently do not have any direct references regarding the use of this product in the application. All of our references come from loyal customers like you, who have used our products, published their findings in journals and later inform us of their publications. Please rest assure that if the product does not perform in species and application as stated on the datasheet, and the problem (not protocol related) has been reported to us within 120 days of purchase, we will be more than happy to offer you a free replacement or refund. Should you require any other assistance, please do not hesitate to contact me.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com