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Human Phospho-CD117/c-kit (Y730) Antibody

R&D Systems, part of Bio-Techne | Catalog # AF3527

R&D Systems, part of Bio-Techne
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AF3527
AF3527-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Simple Western, Western Blot

Cited:

Immunocytochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Product Specifications

Immunogen

Phosphopeptide containing human CD117/c-kit Y730 site

Specificity

Detects human and mouse CD117/c-kit when phosphorylated at Y730.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human Phospho-CD117/c-kit (Y730) Antibody

Detection of Human Phospho-CD117/ c-kit (Y730) antibody by Western Blot.

Detection of Human Phospho-CD117/ c‑kit (Y730) by Western Blot.

Western blot shows lysates of MO7e human megakaryocytic leukemic cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human SCF/c-kit Ligand (Catalog # 255-SC) for 10 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-CD117/c-kit (Y730) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3527), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-CD117/c-kit (Y730) at approximately 145 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human CD117/c-kit antibody by Simple WesternTM.

Detection of Human CD117/c-kit by Simple WesternTM.

Simple Western lane view shows lysates of MO7e human megakaryocytic leukemic cell line, loaded at 0.2 mg/mL. A specific band was detected for CD117/c-kit at approximately 177 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-CD117/c-kit (Y730) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3527) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Applications for Human Phospho-CD117/c-kit (Y730) Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed frozen sections of mouse embryo (E13-15)

Simple Western

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

Western Blot

0.5 µg/mL
Sample: MO7e human megakaryocytic leukemic cell line treated with Recombinant Human SCF (Catalog # 255-SC)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 3 reviews rated 3.7 using AF3527 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

Additional CD117/c-kit Products

Product Documents for Human Phospho-CD117/c-kit (Y730) 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 Phospho-CD117/c-kit (Y730) Antibody

For research use only

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