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Custom Human/Mouse Acetylated-Histone H3 Cell-based ELISA Kit
Custom Human/Mouse Acetylated-Histone H3 Cell-based ELISA Kit
RayBio®Human/Mouse Acetylated-Histone H3 Cell-based ELISA Kit. This ELISA is for measuring Acetylated-Histone H3 in human and mouse adherent cell lines.
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Species | Human, Mouse |
| Specificity | The antibody pair provided in this kit recognizes human/mouse Acetylated-Histone H3. |
| Accession Number | P84243|P84244 |
| Gene Id | 3020|3021 |
| Gene Symbols | H3F3A, H3F3B, H3.3A, H3F3, PP781, H3.3B |
| Protein Name / Synonyms | Histone H3.3 |
| Research Area | Post-Translational Modifications, Phosphorylation, Acetylation, Epigenetics |
| Shipping Type | Blue ice |
| Storage | -20°C |
Product Features
- Rapidly measures acetylated protein in lysates
- Simultaneously measure acetylated 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
- Quneching Buffer
- Blocking Buffer
- Anti-cleaved 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.