JARID1C (KDM5C) Homogeneous Assay Kit
The JARID1C Homogeneous Assay Kit is designed to measure JARID1C activity for screening and profiling applications. JARID1C, also known as SMCX (Smcy homolog, X-linked), KDM5C, or XE169, is a JumonjiC (JmjC) and ARID domain-containing histone lysine demethylase that exhibits demethylation activity toward H3-K4Me2 and H3-K4Me3. The JARID1C Homogeneous Assay Kit comes in a convenient AlphaLISA® format (Scheme 1), with biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified JARID1C for 96 enzyme reactions. The key to the JARID1C Homogeneous Assay Kit is a highly specific antibody that recognizes demethylated substrate. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, a sample containing JARID1C enzyme is incubated with the biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.
Scheme 1: Our histone demethylase assays utilize highly specific antibodies that recognize demethylated histone substrate. First, a sample containing the enzyme is incubated with a biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts, as shown above.
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AlphaLISA® anti-rIgG acceptor beads, 5 mg/ml (PerkinElmer #AL104C)
AlphaScreen® Streptavidin-conjugated donor beads, 5 mg/ml (PerkinElmer #6760002S)
Optiplate -96 (PerkinElmer #6005290) or Optiplate -384 (PerkinElmer #6007290)
AlphaScreen® microplate reader
Adjustable micropipettor and sterile tips
96 reactions:
Catalog # | Component | Amount | Storage | |
50112 | JARID1C (KDM5C) | 20 μg | -80°C | Avoid freeze/ thaw cycles |
52140M4 | Primary antibody 13-4 | 5 µg | -80°C | |
Biotinylated histone H3 peptide substrate (JARID) | 300 reactions | -80°C | ||
52406 | 4x HDM Assay Buffer 1 | 3 ml | -80°C | |
4x Detection buffer D | 2 ml | -20°C |
384 reactions:
Catalog # | Component | Amount | Storage | |
50112 | JARID1C (KDM5C) | 20 μg | -80°C | Avoid freeze/ thaw cycles |
52140M4 | Primary antibody 13-4 | 20 μl | -80°C | |
79840 | Biotinylated histone H3 peptide substrate | 500 reactions | -80°C | |
52406 | 4x HDM Assay Buffer 1 | 3 ml | -80°C | |
4x Detection buffer D | 2 ml | -20°C |