SCF (KITLG) Mouse Monoclonal Antibody

Specifications

Product Data
Applications	ELISA, IF, WB
Recommend Dilution	Sandwich ELISA: In a Sandwich ELISA (assuming 100 µl/well), a concentration of 4.0-8.0 µg/ml of this antibody will detect at least 1000 pg/ml of recombinant human SCF when used with Biotinylated antigen affinity purified anti-Human SCF (Cat.-No PP1066B) as the detection antibody at a concentration of approximately 1.0-2.0 µg/ml.   Western Blot: To detect Human SCF by Western Blot analysis this antibody can be used at a concentration of 0.25-0.50 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant hSCF is 2.0-4.0 ng/lane, under non-reducing conditions.  Immunohistochemistry: This antibody stained CACO-2 cells and A-431 cells. The primary antibody was incubated at 2.0 µg/ml overnight at 4°C followed by a fluorescent labeled secondary antibody. Information and photo are courtesy of the Cell Profiling group, SciLifeLab Stockholm.
Reactivity	Human
Host	Mouse
Clonality	Monoclonal
Immunogen	Highly purified (>98%) E.coli derived Recombinant Human SCF (Cat.-No PA124)
Specificity	Recognizes Human SCF. Other species not tested.
Formulation	PBS without preservatives State: Azide Free State: Lyophilized purified Ig fraction.
Reconstitution Method	Restore in sterile water to a concentration of 1.0 mg/ml.
Concentration	1.0 mg/ml
Purification	Protein A Chromatography
Conjugation	Unconjugated
Storage Condition	Store lyophilized at 2-8°C for 6 months or at -20°C long term. After reconstitution store the antibody undiluted at 2-8°C for one month  or (in aliquots) at -20°C long term. Avoid repeated freezing and thawing.
Gene Name	KIT ligand
Database Link	Entrez Gene 4254 Human UniProt ID P21583
Synonyms	KITL, Kit ligand, c-Kit ligand, Stem cell factor, Mast cell growth factor, MGF
Note	Protocol: Immunofluorescence: General Cell: Please refer to the antibody Product Information Sheet for the primary antibody concentration and the selected cell line. 1. A multiwell plate (Glass bottom, 96-well, 300μL) is coated with fibronectin (conc. 12.5μg/mL) for 1 hour at room temperature (RT).  2. Cells are seeded (10,000-15,000 cells per well) and incubated at 37˚C in humidified air with 5.0% CO2 for at least 4 hours. 3. Growth medium is removed and the cells are washed in PBS (8.1mM Na2HPO4, 1.5mM KH2PO4, 137mM NaCl, 2.7mM KCl, pH 7.2).  4. The cells are fixed for 15 minutes in ice cold 4% paraformaldehyde (pH 7.2-7.3) in growth medium supplemented with 10% fetal bovine serum (FBS).  5. The cells are permeabilized 3 times for 5 minutes each with 0.1% Triton X-100 in PBS.  6. The cells are washed once with PBS. 7. The primary antibody is diluted in PBS supplemented with 4% FBS and incubated overnight at 4°C. 8. The following day, the cells are washed 4 times for 10 minutes each with PBS.  9. The secondary antibody is diluted to 1μg/mL in PBS supplemented with 4% FBS and incubated for 1.5 hours at room temperature.  10. The cells are counterstained for 4 minutes with the nuclear probe DAPI.  11. The cells are washed 4 times for 10 minutes each with PBS before mounted in PBS containing 78% glycerol.
Reference Data
Protein Families	Druggable Genome, Transmembrane
Protein Pathways	Cytokine-cytokine receptor interaction, Hematopoietic cell lineage, Melanogenesis, Pathways in cancer

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