|Product Name||Anti-SCF/KITLG Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Kit ligand/Mast cell growth factor(KITLG) detection. Tested with WB, ELISA in Human.|
|Cite This Product||Anti-SCF/KITLG Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1010)|
|Immunogen||E. coli-derived human SCF recombinant protein(Position: E26-A190).|
Assay Dilutions Overview
ELISA , 0.1-0.5μg/ml, Human
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
Images And Assay Conditions
Figure. Western blot analysis of SCF using anti- SCF antibody (RP1010).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane : Recombinant Human SCF Protein 0.5ng,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- SCF antigen affinity purified polyclonal antibody (Catalog # RP1010) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SCF at approximately 18KD. The expected band size for SCF is at 18KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Kit ligand|
|Alternative Names||Kit ligand;Mast cell growth factor;MGF;Stem cell factor;SCF;c-Kit ligand;Soluble KIT ligand;sKITLG;KITLG;MGF, SCF;|
|Subcellular Localization||Isoform 1: Cell membrane; Single-pass type I membrane protein.|
|Molecular Weight||30899 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein 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.|
|Background||Stem Cell Factor(also known as SCF, kit-ligand, KL, or steel factor) is a cytokine that binds to the c-Kit receptor(CD117). The SCF gene is mapped to 12q21.32. SCF can exist both as a transmembrane protein and a soluble protein. This cytokine plays an important role in hematopoiesis(formation of blood cells), spermatogenesis, and melanogenesis. SCF may be used along with other cytokines to culture HSCs and hematopoietic progenitors. The expansion of these cells ex-vivo(outside the body) would allow advances in bone-marrow transplantation, in which HSCs are transferred to a patient to re-establish blood formation. One of the problems of injecting SCF for therapeutic purposes is that SCF activates mast cells. The injection of SCF has been shown to cause allergic-like symptoms and the proliferation of mast cells and melanocytes.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,