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Human CD117/c-kit Antibody

R&D Systems, part of Bio-Techne | Catalog # AF332

R&D Systems, part of Bio-Techne
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AF332
AF332-SP

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

CyTOF-ready, Flow Cytometry, Immunohistochemistry, Neutralization, Simple Western, Western Blot

Cited:

Immunocytochemistry, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CD117/c-kit
Gln26-Thr520
Accession # P10721

Specificity

Detects human and mouse CD117/c-kit in direct ELISAs and Western blots. In Western blots, approximately 2% cross-reactivity with recombinant human PDGF sR alpha is observed.

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 CD117/c-kit Antibody

Cell Proliferation Induced by SCF/c‑kit Ligand and Neutralization by Human CD117/c-kit Antibody.

Cell Proliferation Induced by SCF/c‑kit Ligand and Neutralization by Human CD117/c-kit Antibody.

Recombinant Human SCF/c-kit Ligand (Catalog # 255-SC) induces cell proliferation in the TF-1 human erythroleukemic cell line in a dose-dependent manner (orange line), as measured by the Resazurin (Catalog # AR002). Under these conditions, proliferation elicited by SCF/c-kit Ligand is neutralized (green line) by increasing concentrations of Goat Anti-Human CD117/c-kit Antigen Affinity-purified Polyclonal Antibody (Catalog # AF332). The ND50 is typically 0.06-0.36 µg/mL in the presence of 20 ng/mL of Recombinant Human SCF/c-kit Ligand.
CD117/c-kit antibody in Human Brain by Immunohistochemistry (IHC-P).

CD117/c-kit in Human Brain.

CD117/c-kit was detected in immersion fixed paraffin-embedded sections of human brain (hippocampus) using Goat Anti-Human CD117/c-kit Antigen Affinity-purified Polyclonal Antibody (Catalog # AF332) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human CD117/c-kit antibody by Western Blot.

Detection of Human CD117/c-kit by Western Blot.

Western blot shows lysates of MO7e human megakaryocytic leukemic cell line. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human CD117/c-kit Antigen Affinity-purified Polyclonal Antibody (Catalog # AF332) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CD117/c-kit at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Applications for Human CD117/c-kit Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: TF-1 human erythroleukemic cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (hippocampus) subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Simple Western

5 µg/mL
Sample: MO7e human megakaryocytic leukemic cell line

Western Blot

0.1 µg/mL
Sample: MO7e human megakaryocytic leukemic cell line

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 0.06-0.36 μg/mL in the presence of 20 ng/mL Recombinant Human SCF/c‑kit Ligand.

Reviewed Applications

Read 2 reviews rated 4.5 using AF332 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.

Background: CD117/c-kit

Stem cell factor receptor (CD117, the gene product of the c-kit proto-oncogene) and its ligand, stem cell factor (also named c-kit ligand, mast cell growth factor), play essential roles in gametogenesis, melanogenesis and hematopoiesis. The human stem cell factor receptor cDNA encodes a 972 amino acid (aa) residue precursor membrane protein with a 25 aa residue signal peptide (experimentally determined), a 495 aa residue extracellular domain, a 23 aa residue transmembrane segment and a 429 aa residue cytoplasmic domain. Stem cell factor receptor is a member of the type III subfamily of receptor tyrosine kinases (RTK) that also includes the receptors for M-CSF, Flt-3, PDGF, and VEGF. All class III RTKs are characterized by the presence of five immunoglobulin-like domains in their extracellular region and a split kinase domain in their intracellular region. SCF binding induces receptor homodimerization and signal transduction. SCF receptor is expressed in hematopoietic progenitor cells, normal B- and T-lymphocyte progenitor cells, mast cells, germ cells, melanocytes, neurons, glial cells, placenta, kidney, lung, and gut. In addition, SCF receptor expression has also been reported in a number of human tumor cell lines. SCF receptor can be proteolytically cleaved from the cell surface and high levels of soluble SCF receptor has been detected in cell conditioned medium and human plasma. Recombinant soluble SCF receptor binds SCF with high affinity and is a potent SCF antagonist.

References

  1. Broudy, V. (1997) Blood 90:1345.
  2. Vliagoftis, H. et al. (1997) J. Allergy Clin. Immunol. 100:435.

Long Name

Stem Cell Factor Receptor

Alternate Names

c-kit, CD117, ckit, KIT, PBT, SCF R, SCFR

Entrez Gene IDs

3815 (Human); 16590 (Mouse); 64030 (Rat)

Gene Symbol

KIT

UniProt

Additional CD117/c-kit Products

Product Documents for Human CD117/c-kit Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human CD117/c-kit Antibody

For research use only

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