RayBiotech Human Phospho-FLT3 (Y589) and Total FLT3 ELISA
Phosphorylation ELISA. Catalog #: PEL-FLT3-Y589-T
Human Phospho-FLT3 (Y589) and Total FLT3 ELISA
RayBio® Human Phospho-FLT3 (Tyr589) and Total FLT3 ELISA Kit. This assay semi-quantitatively measures FLT3 phosphorylated at Tyrosine-589 as well as total FLT3 in cell lysate samples.
Specifications
Size |
1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
---|---|
Species |
Human |
Quantitative/Semi-Quantitative |
Semi-Quantitative |
Protein Name / Synonyms |
Receptor-type tyrosine-protein kinase FLT3 (EC 2.7.10.1) (FL cytokine receptor) (Fetal liver kinase-2) (FLK-2) (Fms-like tyrosine kinase 3) (FLT-3) (Stem cell tyrosine kinase 1) (STK-1) (CD antigen CD135) |
Specificity |
The antibody pair provided in this kit recognizes Human FLT3 phosphorylated at site Tyrosine-589 as well as total FLT3 |
Accession Number |
P36888 |
Gene Id |
2322 |
Compatible Sample Types |
Tissue Lysates, Cell Lysates |
Solid Support |
96-well Microplate |
Method Of Detection |
Colorimetric |
Design Principle |
Sandwich-based |
Research Area |
Post-Translational Modifications, Phosphorylation, Tyrosine Kinase Family |
Product Features
- Rapidly measure phosphorylated protein in lysates
- Screen numerous different cell lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Anti-Phospho Antibody
- HRP-Conjugated Secondary Antibody
- Assay Diluent
- TMB One-Step Substrate
- Stop Solution
- Lysis Buffer
- Positive Control Sample
Other Materials Required
- Distilled or deionized water
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare sample dilutions
- Protease and Phosphatase inhibitors
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- Benchtop rocker or shaker
- Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
1. Prepare all reagents and samples as instructed in the manual.
2. Add 100 µl of sample or positive control to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 µl of prepared primary antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 µl of prepared 1X HRP-Streptavidin to each well.
7. Incubate 1 h at RT.
8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µl of Stop Solution to each well.
11. Read at 450 nm immediately.
Typical Data
Positive Control
Jurkat cells were treated with Pervanadate. Solubilize cells at 4 x 10^7 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
Pervanadate Stimulation of A431 Cell Line
A431 cells were treated or untreated with Pervanadate. Cell lysates were analyzed using this phosphoELISA and Western Blot.
Storage/Stability
Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.
Catalog #: PEL-FLT3-Y589-T-1 -- 1 Plate Kit
Catalog #: PEL-FLT3-Y589-T-2 -- 2 Plate Kit
Catalog #: PEL-FLT3-Y589-T-5 -- 5 Plate Kit
Please email us at cs@medikonia.com for any enquiry. To place an order, please include the catalog number(s) of the product(s) in the email.