Human Phospho-Flt-3/Flk-2 (Y591) Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Flt-3 when phosphorylated at Y591.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Phosphopeptide containing human Flt-3 Y591 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples

Detection of Human Phospho-Flt‑3/Flk‑2 (Y591) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated
(-) or treated (+) with 100 μM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-Flt‑3/Flk‑2 (Y591) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF368), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-Flt‑3/Flk‑2 (Y591) at approximately 160 and 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

Phospho-Flt‑3/Flk‑2 (Y591) in A431 Human Cell Line. Flt‑3/Flk‑2 phosphorylated at Y591 was detected in immersion fixed A431 human epithelial carcinoma cell line untreated (lower panel) or treated (upper panel) with pervanadate using Rabbit Anti-Human Phospho-Flt‑3/Flk‑2 (Y591) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF368) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Phospho-Flt‑3/Flk‑2 (Y591) by Simple WesternTM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 µM Pervanadate (PV) for 10 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Flt‑3/Flk‑2 (Y591) at approximately 166 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-Flt‑3/Flk‑2 (Y591) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF368). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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: Flt-3/Flk-2

The Flt-3 (fms-like tyrosine kinase) receptor, also named Flk-2 (fetal liver kinase) and Stk-1(stem cell tyrosine kinase) is a member of the class III subfamily of receptor tyrosine kinases that also includes KIT, the receptor for SCF and FMS, the receptor for M-CSF. The extracellular region of these receptors contains five immunoglobulin-like domains and the intracellular region contains a split kinase domain. Human Flt-3 cDNA encodes a 993 amino acid (aa) residue type I membrane protein with a 26 aa residue signal peptide, a 515 aa extracellular domain with 10 potential N-linked glycosylation sites, a 21 aa residue transmembrane domain and a 431 aa residue cytoplasmic domain. Mouse Flt-3 has also been cloned and shown to share 85% amino acid sequence identity with human Flt-3. Flt-3 expression has been detected in various tissues, including placenta, gonads, and tissues of nervous and hematopoietic origin. Among hematopoietic cells, the expression of Flt-3 was found to be restricted to the highly enriched stem/progenitor cell populations. The ligand for Flt-3 (FL) has been identified to be a transmembrane protein with structural homology to M-CSF and SCF. Recombinant soluble Flt-3/Fc chimeric protein has been shown to bind FL with high affinity and is a potent FL antagonist.

  • References:
    1. Rosnet, O. et al. (1996) Acta. Haemato. 95:218.
    2. Drexler, H.G. (1996) Leukemia 10:588.
  • Long Name:
    fms-like Tyrosine Kinase 3
  • Entrez Gene IDs:
    2322 (Human); 14255 (Mouse)
  • Alternate Names:
    CD135 antigen; CD135; EC 2.7.10; fetal liver kinase 2; FL cytokine receptor; FLK2; Flk-2; FLT3 receptor tyrosine kinase; Flt3; Flt-3; Fms-like tyrosine kinase 3; fms-related tyrosine kinase 3; growth factor receptor tyrosine kinase type III; Stem cell tyrosine kinase 1; STK-1; STK1EC 2.7.10.1; Tyrosine-protein kinase receptor FLT3
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