FBXL11 (KDM2A) Homogeneous Assay Kit
The FBXL11 (KDM2A) Homogeneous Assay Kit is designed to measure FBXL11 (F-box and leucine-rich repeat protein 11) activity for screening and profiling applications. The FBXL11 (KDM2A) Homogeneous Assay Kit comes in a convenient 96-well or 384-well AlphaLISA® format, with enough purified recombinant FBXL11 (amino acids 2-700), substrate, primary antibody, assay and detection buffer for 100 or 400 enzyme reactions.
Figure 1: FBXL11 (KDM2A) Homogenous Assay Kit schematic.
A sample containing FBXL11 is incubated with a biotinylated substrate. This is followed by the addition of acceptor beads and primary antibody, and finally donor beads. Alpha-counts are then counted. Alpha-counts are proportional to FBXL11 demethylase activity
- AlphaLISA® Anti-Rabbit IgG Acceptor Beads, 5 mg/ml (Perkin Elmer #AL104C)
- AlphaScreen® Streptavidin-Conjugated Donor Beads, 5 mg/ml (Perkin Elmer #6760002S)
- Optiplate - 96 (Perkin Elmer #6005290)
- AlphaScreen® microplate reader
96 reactions
| Catalog # | Name | Amount | Storage |
| 50156 | FBXL11 (KDM2A, JHDM1A), FLAG-Avi-tags* | 10 µg | -80°C |
| 52140P-2 | Primary Antibody 16-2 | 5 µl | -80°C |
| 79843 | Biotinylated Histone H3 Peptide Substrate | 300 reactions | -80°C |
| 79847 | 4x HDM Assay Buffer 5 | 3 ml | -80°C |
| 52301 | 4x Detection Buffer | 2 ml | -20°C |
*The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.
384 reactions
| Catalog # | Name | Amount | Storage |
| 50156 | FBXL11 (KDM2A, JHDM1A), FLAG-Avi-tags* | 20 µg | -80°C |
| 52140P-2 | Primary Antibody 16-2 | 20 µl | -80°C |
| 79843 | Biotinylated Histone H3 Peptide Substrate | 500 reactions | -80°C |
| 7984 | 4x HDM Assay Buffer 5 | 3 ml | -80°C |
| 52301 | 4x Detection Buffer | 2 ml | -20°C |
*The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.
FBXL11 (F-box and leucine-rich repeat protein 11), also known as KMD2A (lysine demethylase 2A), belongs to the JmjC (Jumonji C domain) demethylase family of proteins. It is involved in H3 lysine 36 (H3K36) demethylation, and chromosome remodeling and gene expression, stem cell differentiation, metabolism, and DDR (DNA damage repair). It is found in the nucleus, where it can bind to cpG DNA, and it is found at high levels in the brain, testis, ovaries, and the lungs. It can also be found in many cancer types, with the notable exception of prostate cancer. FBXL11 expression is regulated by microRNA (miRNA), and its levels increase during inflammation, hypoxia or oxidative stress. For instance, in non-small cell lung cancer (NSCLC) tPA (tissue-type plasminogen activator) can activate COX2 (cyclooxygenase-2) by acting on FBXL11. FBXL11 promotes NSCLC also by binding to the DUSP3 (dual specificity phosphatase 3) promoter region, decreasing its expression, and consequently increasing phosphorylation of ERK1/2 (extracellular signal regulated kinases 1/2) and cancer metastasis. The use of FBXL11 as a therapeutic target is thus an interesting approach and is under investigation.
Iwase S., et al.,2007 Cell 128(6):1077-1088.
Liu L., et al., 2021 Exp Ther Med 22(1):723.
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