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蚂蚁淘在线 / 品牌中心 / RayBiotech / RayBiotech/Custom Human Phospho-ATF2 (Thr69/71) and Total ATF2 Cell-Based ELISA/1 Plate Kit/CBEL-ATF2-T69-1
商品描述
Antigen Information
Accession Number:
- P15336
- P16951
Gene ID:
- 1386
Gene Symbols:
- ATF2
- CREB2
- CREBP1
Protein Name & Synonyms:
Cyclic AMP-dependent transcription factor ATF-2 (cAMP-dependent transcription factor ATF-2) (EC 2.3.1.48) (Activating transcription factor 2) (Cyclic AMP-responsive element-binding protein 2) (CREB-2) (cAMP-responsive element-binding protein 2) (HB16) (Histone acetyltransferase ATF2) (cAMP response element-binding protein CRE-BP1)
Target Species:
- Human
Assay Format
Pathway:
- MAPK Signaling
- PI3K-AKT Signaling
- TGF-beta Signaling
Specificity:
This ELISA kit recognizes Human ATF2 phosphorylated at site Threonine-69/71 as well as total ATF2.
Number of Targets Detected:
2
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 Strip Microplate Kit
Product Features
- Rapidly measure phosphorylated protein in adherent cell lines
- Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
- No sample lysis is needed
- Compatible with a standard ELISA plate reader
Application Notes
Kit Components
- Uncoated 96-well Strip Microplate
- Wash Buffers
- Fixing Solution
- Quenching Buffer
- Blocking Buffer
- Anti-phospho antibody
- Anti-pan antibody
- HRP-Conjugated Secondary Antibody
- TMB One-Step Substrate
- Stop Solution
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 manufacturer"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
Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.
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