Human SCF/c‑kit Ligand Antibody

R&D Systems | Catalog # AB-255-NA

R&D Systems
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Neutralization

Cited:

Western Blot, Neutralization, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all SCF/c-kit Ligand Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human SCF/c‑kit
Glu26-Ala189
Accession # P21583

Specificity

Detects human SCF/c‑kit Ligand in direct ELISAs and Western blots. In direct ELISA, approximately 45% cross‑reactivity with recombinant mouse SCF is observed, approximately 30% cross-reactivity with recombinant canine SCF, recombinant rat SCF, and recombinant feline SCF is observed. It will also neutralize the biological activity of recombinant mouse SCF at a 10 fold higher IgG concentration.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human SCF/c‑kit Ligand Antibody

Cell Proliferation Induced by SCF/c‑kit Ligand and Neutralization by Human SCF/c‑kit Ligand Antibody.

Cell Proliferation Induced by SCF/c‑kit Ligand and Neutralization by Human SCF/c‑kit Ligand Antibody.

Recombinant Human SCF/c-kit Ligand (Catalog # 255-SC) stimulates proliferation in the TF-1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human SCF/c-kit Ligand (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human SCF/c-kit Ligand Polyclonal Antibody (Catalog # AB-255-NA). The ND50 is typically 0.4-2 µg/mL.

Detection of SCF/c-kit Ligand by Western Blot

Detection of SCF/c-kit Ligand by Western Blot

IGF1 and GSK3i stimulate Kitl/KITLG transcription. Results obtained in murine and human smooth muscle cells (A-E), human LX-2 stellate cells (F-G) and human GIST-T1 cells (H-I) are shown. A, Hoffman modulation contrast image of primary human gastric smooth muscle cells. B, KITLG mRNA (total: soluble+membrane-bound) was readily detectable in primary human smooth muscle cells (passage 3) maintained with Smooth Muscle Growth Medium-2 containing insulin, hFGF-B, hEGF and 5% FBS (Lonza) but not in 24-h growth factor- and serum-deficient basal medium. C, Both IGF1 (100 ng/mL) and the GSK3 alpha / beta inhibitor SB415286 (30 µM) stimulated Kitl expression in murine gastric tunicamuscularis organotypic cultures (n=3/group). D-E, SB415286 stimulated endogenous Kitl expression in murine primary gastric smooth muscle cells (D; n=3/group) and KITLG transcriptional activity in the same cell type transfected with a KITLG promoter (-2120 bp to +407 bp)-pGL3 luciferase construct (E; n=3/group). IGF1 (100 ng/mL; n=3/group) and SB415286 (30 µM; n=6/group) also increased endogenous KITLG mRNA expression in LX-2 (F) and GIST-T1 cells (H) and stimulated KITLG promoter activity in a time-dependent fashion (LX-2: n=6-9/group; G; GIST-T1: n=3-11/group; I). Groups marked by asterisk are different from the control group, and groups not sharing the same superscript letter are different from each other (P<0.05 by post-hoc multiple comparisons). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24116170), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human SCF/c‑kit Ligand Antibody

Application
Recommended Usage

Western Blot

1 µg/mL
Sample: Recombinant Human SCF/c-kit Ligand (Catalog # 255-SC)

Neutralization

Measured by its ability to neutralize SCF/c‑kit Ligand-induced proliferation in the TF‑1 human erythroleukemic cell line. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323. The Neutralization Dose (ND50) is typically 0.4-2 µg/mL in the presence of 10 ng/mL Recombinant Human SCF/c‑kit Ligand.

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Reconstitution

Reconstitute at 1 mg/mL in sterile PBS.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: SCF/c-kit Ligand

Stem cell factor (SCF), also known as c-kit ligand (KL), mast cell growth factor (MGF), and steel factor (SLF), is a widely expressed 28‑40 kDa type I transmembrane glycoprotein (1). It promotes the survival, differentiation, and mobilization of multiple cell types including myeloid, erythroid, megakaryocytic, lymphoid, germ cell, and melanocyte progenitors (1‑7). SCF is a primary growth and activation factor for mast cells and eosinophils (8). Mature human SCF consists of a 189 amino acid (aa) extracellular domain (ECD), a 23 aa transmembrane segment, and a 36 aa cytoplasmic tail (9). The ECD shows both N-linked and O-linked glycosylation (10). Proteolytic cleavage at two alternate sites in the extracellular juxtamembrane region releases a 25 kDa soluble molecule which is comparable to the only form produced by Steel-dickie mutant mice (11, 12). An alternately spliced isoform of human SCF lacks 28 aa that encompasses the primary proteolytic recognition site (13). Within the ECD of the short isoform (corresponding to this recombinant protein), human SCF shares 75%‑83% aa sequence identity with canine, feline, mouse, and rat SCF. Rat SCF is active on mouse and human cells, but human SCF is only weakly active on mouse cells (9). Noncovalent dimers of transmembrane or soluble SCF interact with the receptor tyrosine kinase SCF R/c-kit to trigger receptor dimerization and signaling (14). SCF assists in the recovery of cardiac function following myocardial infarction by increasing the number of cardiomyocytes and vascular channels (15).

References

  1. Ashman, L.K. (1999) Int. J. Biochem. Cell Biol. 31:1037. 
  2. Sette, C. et al. (2000) Int. J. Dev. Biol. 44:599. 
  3. Yoshida, H. et al. (2001) J. Invest. Dermatol. Symp. Proc. 6:1. 
  4. Erlandsson, A. et al. (2004) Exp. Cell Res. 301:201. 
  5. Kapur, R. et al. (2002) Blood 100:1287. 
  6. Wang, C.-H. et al. (2007) Arterioscler. Thromb. Vasc. Biol. 27:540. 
  7. Bashamboo, A. et al. (2006) J. Cell Sci. 119:3039. 
  8. Reber, L. et al. (2006) Eur. J. Pharmacol. 533:327.
  9. Martin, F.H. et al. (1990) Cell 63:203.
  10. Arakawa, T. et al. (1991) J. Biol. Chem. 266:18942.
  11. Majumdar, M.K. et al. (1994) J. Biol. Chem. 269:1237.
  12. Brannan, C.I. et al. (1991) Proc. Natl. Acad. Sci. 88:4671.
  13. Anderson, D.M. et al. (1991) Cell Growth Differ. 2:373.
  14. Lemmon, M.A. et al. (1997) J. Biol. Chem. 272:6311.
  15. Kanellakis, P. et al. (2006) Cardiovasc. Res. 70:117.

Long Name

Stem Cell Factor

Alternate Names

c-kit Ligand, DCUA, DFNA69, FPH2, FPHH, KITLG, KL-1, MGF, SHEP7, SLF

Entrez Gene IDs

4254 (Human); 17311 (Mouse); 60427 (Rat); 403507 (Canine); 493937 (Feline)

Gene Symbol

KITLG

UniProt

P21583

Additional SCF/c-kit Ligand Products

Product Documents for Human SCF/c‑kit Ligand Antibody

Certificate of Analysis

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Product Specific Notices for Human SCF/c‑kit Ligand Antibody

For research use only

Citations for Human SCF/c‑kit Ligand Antibody

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