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

Catalog # Availability Size / Price Qty
AF368
AF368-SP
Detection of Human Phospho-Flt‑3/Flk‑2 (Y591) by Western Blot.
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Human Phospho-Flt-3/Flk-2 (Y591) Antibody Summary

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

Western Blot Detection of Human Phospho-Flt‑3/Flk‑2 (Y591) by Western Blot. View Larger

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. View Larger

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.

Simple Western Detection of Human Phospho-Flt‑3/Flk‑2 (Y591) by Simple WesternTM. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

FAQs

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Cell and Tissue Staining Kits

Isotype Controls

Normal Rabbit IgG Control

Ctrl,CyTOF-ready

Reconstitution Buffers

Secondary Antibodies

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