The RayBio® Sumoylated-HDAC1 and Total-HDAC1 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human and mouse sumoylated histone deacetylase 1 and total histone deacetylase 1. An anti-pan HDAC1 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and HDAC1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-sumoylation antibody is used to detect sumoylated HDAC1 or biotinylated anti-HDAC1 antibody is used to detect total HDAC1. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of sumoylated HDAC1 or total HDAC1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

• Quantify quickly and easily
• Screen numerous different lysates without performing a Western Blot analysis
• Minimal hands-on time, convenient, and non-radioactive material

• Microplate reader capable of measuring absorbance at 450 nm
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Log-log graph paper or computer and software for ELISA data analysis
• Absorbent paper
• Tubes to prepare positive control or sample dilutions
• Distilled or deionized water
• Shaker

1. Prepare all reagents, samples and Positive Control as instructed.
2. Add 100 μLsample and Positive Control to each well. Incubate 2.5 hours at room temperature or overnight at 4ºC with gentle shaking.
3. Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
4. Add 100 μL prepared HRP-Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.