Protein Name & Synonyms:
Tubulin alpha-1A chain (Alpha-tubulin 3)(Tubulin B-alpha-1)(Tubulin alpha-3 chain)
The antibody pair provided in this kit recognizes Human
Number of Targets Detected:
1, 2, or 5 x 96-Well Strip Microplate Kit
RayBio® Acetylated-Alpha Tubulin and Alpha Tubulin ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human, mouse and rat cell lysates. By determining Acetylated Alpha Tubulin protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
- Rapidly measures acetylated protein in lysates
- Screen numerous different cell lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Assay Diluent
- Anti-Acetylated Antibody
- Anti-Pan Antibody
- HRP-Conjugated Secondary Antibody
- 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
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared 1X HRP-Streptavidin to each well.
- Incubate 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.
COS cells were treated with Trichostatin A (TSA). Solubilize cells at 4 x 107
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.
TSA Stimulation of NIH/3T3 Cell Line
NIH/3T3 cells were treated or untreated with TSA. Cell lysates were analyzed using this ELISA and Western Blot.
Upon receipt, the kit should be stored at â20°C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4°C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge, and store at â20°C. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C.