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.
Human Phospho-Flt‑3/Flk‑2 (Y591) Antibody
R&D Systems | Catalog # AF368
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Key Product Details
Validated by
Biological Validation
Species Reactivity
Human
Applications
Western Blot, Immunocytochemistry, Simple Western
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
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Product Specifications
Immunogen
Phosphopeptide containing human Flt-3 Y591 site
Specificity
Detects human Flt-3 when phosphorylated at Y591.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human Phospho-Flt‑3/Flk‑2 (Y591) Antibody
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.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.Applications for Human Phospho-Flt‑3/Flk‑2 (Y591) Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed A431 human epithelial carcinoma cell line treated with recombinant human EGF (Catalog # 236-EG-01M), and immersion fixed A431 human epithelial carcinoma cell line treated with pervanadate
Sample: Immersion fixed A431 human epithelial carcinoma cell line treated with recombinant human EGF (Catalog # 236-EG-01M), and immersion fixed A431 human epithelial carcinoma cell line treated with pervanadate
Simple Western
5 µg/mL
Sample: A431 human epithelial carcinoma cell line treated with Pervanadate (PV)
Sample: A431 human epithelial carcinoma cell line treated with Pervanadate (PV)
Western Blot
0.5 µg/mL
Sample: Pervanadate-treated A431 human epithelial carcinoma cell line
Sample: Pervanadate-treated A431 human epithelial carcinoma cell line
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.
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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.
Calculators
Background: Flt-3/Flk-2
References
- Rosnet, O. et al. (1996) Acta. Haemato. 95:218.
- Drexler, H.G. (1996) Leukemia 10:588.
Long Name
fms-like Tyrosine Kinase 3
Alternate Names
CD135, Flk-2, Flt3, STK-1
Gene Symbol
FLT3
Additional Flt-3/Flk-2 Products
Product Documents for Human Phospho-Flt‑3/Flk‑2 (Y591) Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Phospho-Flt‑3/Flk‑2 (Y591) Antibody
For research use only
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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