Anti-SCF antibody (ab64677)
Key features and details
- Rabbit polyclonal to SCF
- Suitable for: IHC-P, ICC/IF, WB
- Reacts with: Human, African green monkey, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-SCF antibody
See all SCF primary antibodies -
Description
Rabbit polyclonal to SCF -
Host species
Rabbit -
Specificity
Recent retest of the latest lot of this antibody have shown that this product cannot be longer guaranteed in WB using cell lines and tissue lysates expressing the endogenous protein only. Instead batches have passed in WB using recombinant SCF lysate. All batches are still tested successfully in both ICC and IHC (endogenous protein). (Sept 2017) -
Tested applications
Suitable for: IHC-P, ICC/IF, WBmore details -
Species reactivity
Reacts with: Human, African green monkey, Recombinant fragment
Predicted to work with: Mouse, Rat, Sheep, Goat, Horse, Cow, Cat, Dog, Pig, Monkey, Rhesus monkey
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human SCF.
Read Abcam's proprietary immunogen policy (Peptide available as ab73180.) -
Positive control
- This antibody gave a positive signal in recombinant SCF protein. ICC/IF: HepG2 cells. IHC: Human normal colon
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab64677 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 IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. ICC/IF Use a concentration of 1 - 5 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 31 kDa). Target
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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
modificationsA 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
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Database links
- Entrez Gene: 493937 Cat
- Entrez Gene: 403507 Dog
- Entrez Gene: 4254 Human
- Entrez Gene: 17311 Mouse
- Entrez Gene: 60427 Rat
- Omim: 184745 Human
- SwissProt: P79169 Cat
- SwissProt: Q06220 Dog
see all -
Alternative names
- C kit ligand antibody
- C-kit ligand antibody
- Ckit ligand antibody
see all
Images
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Western blot - Anti-SCF antibody (ab64677)Anti-SCF antibody (ab64677) at 1 µg/ml + SCF Recombinant Protein at 0.1 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab64677 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Immunocytochemistry/ Immunofluorescence - Anti-SCF antibody (ab64677)ab64677 stained in HepG2 cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab64677 at 5 µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCF antibody (ab64677)IHC image of SCF antibody staining in a section of formalin-fixed paraffin-embedded normal human colon* performed on a Leica BONDTM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab64677, 0.1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Protocols
References (15)
ab64677 has been referenced in 15 publications.
- Komatsu K & Masubuchi S Increased supply from blood vessels promotes the activation of dormant primordial follicles in mouse ovaries. J Reprod Dev 66:105-113 (2020). PubMed: 31902808
- Xu M et al. Chronic restraint stress induces excessive activation of primordial follicles in mice ovaries. PLoS One 13:e0194894 (2018). PubMed: 29601583
- Ren K et al. TNF-a inhibits SCF, ghrelin, and substance P expressions through the NF-?B pathway activation in interstitial cells of Cajal. Braz J Med Biol Res 51:e7065 (2018). PubMed: 29694505
- Kobayashi M et al. Bioengineering functional smooth muscle with spontaneous rhythmic contraction in vitro. Sci Rep 8:13544 (2018). PubMed: 30202095
- Giglio D et al. Changes in the Neuronal Control of the Urinary Bladder in a Model of Radiation Cystitis. J Pharmacol Exp Ther 365:327-335 (2018). PubMed: 29530925
Images
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Western blot - Anti-SCF antibody (ab64677)Anti-SCF antibody (ab64677) at 1 µg/ml + SCF Recombinant Protein at 0.1 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab64677 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Immunocytochemistry/ Immunofluorescence - Anti-SCF antibody (ab64677)
ab64677 stained in HepG2 cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab64677 at 5 µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCF antibody (ab64677)
IHC image of SCF antibody staining in a section of formalin-fixed paraffin-embedded normal human colon* performed on a Leica BONDTM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab64677, 0.1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
