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| CODE | Number of Plates | Price | ||
|---|---|---|---|---|
|
CBEL-Tyrosine-2
|
$567.00 | ||
|
|
CBEL-Tyrosine-5
|
$1,260.00 | ||
|
|
CBEL-Tyrosine-1
|
$315.00 |
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Assay Format
Pathway:
- Tyrosine Kinase Family
Specificity:
The antibodies provided in this kit recognizes human, mouse and rat Phosphotyrosine.
Number of Targets Detected:
1
Species Detected:
- Human
Compatible Sample Types:
- Adherent Cell Culture
Design Principle:
- Cell-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
- Semi-Quantitative
Solid Support:
96-well Microplate
Product Specifications
Size:
1, 2, or 5 x 96-Well Microplate Kit
Product Features
- Site and signal pathway-specific
- In vitro detection of adherent cell culture
- No sample lysis needed
- Compatible with a standard ELISA plate reader
- Faster results than with ELISA
- Adaptable for high-throughput screening and drug discovery
Application Notes
Kit Components
- Cell Culture Microplate
- Wash Buffer A
- Wash Buffer B
- Fixing Solution
- HRP-Conjugated Secondary Antibody
- Blocking Buffer
- TMB One-Step Substrate
- Stop Solution
- Quenching Buffer
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
- Seed 10,000-30,000 cells into each well and incubate overnight.
- Apply various treatment, inhibitors or activators according to manufacture's instructions.
- Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
- Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
- Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C.
- Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
- Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Storage/Stability
-20°C
-
Banerjee, Aditi, et al. "Unfolded protein response is required in nu/nu mice microvasculature for treating breast tumor with tunicamycin." Journal of Biological Chemistry 286.33 (2011): 29127-29138.
Species: Human
Sample type: Cell Lysate -
Welters HJ., et al. The protein tyrosine phosphatase-BL, modulates pancreatic ?-cell proliferation by interaction with the Wnt signalling pathway. J Endocrinol. 2008 Jun;197(3):543-52. doi: 10.1677/JOE-07-0262.
Species: Human
Sample type: Cell Lysate -
Kanangat S, Postlethwaite A, Hasty K, Kang A, et al. Induction of multiple matrix metalloproteinases in human dermal and synovial fibroblasts by Staphylococcus aureus: Implications in the pathogenesis of septic arthritis and other soft tissue infections. Arthritis Res Ther. 2006;8:R176. Available at http://arthritis-research.com/content/8/6/R176. Last accessed January 17, 2008.
Species: Human
Sample type: Conditioned Media
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Usage
This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.
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