| Product Includes | Product # | Quantity | Color |
|---|---|---|---|
| FastScan™ ELISA Microwell Strip Plate, 96 Well | 53257 | 96 tests | |
| Histone H3 Rabbit Capture mAb | 10606 | 1 ea | Green (Lyophilized) |
| Citrullinated Histone H3 (Arg17) Rabbit HRP-linked mAb | 28967 | 1 ea | Red (Lyophilized) |
| FastScan™ ELISA Capture Antibody Diluent | 16076 | 3 ml | Green |
| FastScan™ ELISA HRP Antibody Diluent | 28120 | 3 ml | |
| TMB Substrate | 7004 | 11 ml | |
| STOP Solution | 7002 | 11 ml | |
| Sealing Tape | 54503 | 1 ea | |
| ELISA Wash Buffer (20X) | 9801 | 25 ml | |
| FastScan™ ELISA Cell Extraction Buffer (5X) | 69905 | 10 ml | |
| FastScan™ ELISA Cell Extraction Enhancer Solution (50X) | 25243 | 1 ml | |
| FastScan™ ELISA Kit #82818 Positive Control Type 2 | 96313 | 1 ea |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The FastScan™ Citrullinated Histone H3 (Arg17) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of histone H3 when citrullinated at Arg17. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with citrullinated histone H3 (Arg17) in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of citrullinated histone H3 (Arg17).
*Antibodies in kit are custom formulations specific to kit.
IMPORTANT: This FastScan™ ELISA Kit requires 4 washes at Step 6 of the protocol.
Specificity/Sensitivity
Background
The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Histone posttranslational modifications, such as methylation, acetylation, ubiquitination, and citrullination, influence chromatin accessibility and regulate gene expression (1). Histone citrullination is carried out by protein arginine deiminases (PADs), which convert positively charged arginine residues into neutral citrulline (2). Histone citrullination affects the binding of histone reader proteins to chromatin, and it is associated with both transcriptional activation and repression (3-5). Histone citrullination is also a major marker for NETosis, which is a programmed cell death mechanism carried out by neutrophils (6). De-regulation of NETosis is associated with multiple autoimmune diseases, including rheumatoid arthritis (RA) and lupus (7,8). Histone hypercitrullination via PAD4 may also play a role in the progression of various cancers, including multiple myeloma (MM), acute myeloid leukemia (AML), lung cancer, and hepatocellular carcinoma (HCC) (9-12).
- Shanmugam, M.K. et al. (2018) Oncotarget 9, 11414-11426.
- Fuhrmann, J. and Thompson, P.R. (2016) ACS Chem Biol 11, 654-68.
- Wang, Y. et al. (2004) Science 306, 279-83.
- Cuthbert, G.L. et al. (2004) Cell 118, 545-53.
- Zhang, X. et al. (2012) Proc Natl Acad Sci U S A 109, 13331-6.
- Hamam, H.J. and Palaniyar, N. (2019) Biomolecules 9, 369. doi: 10.3390/biom9080369.
- Corsiero, E. et al. (2016) Front Immunol 7, 485.
- Hakkim, A. et al. (2010) Proc Natl Acad Sci U S A 107, 9813-8.
- McNee, G. et al. (2017) Leukemia 31, 373-381.
- Song, G. et al. (2016) Oncotarget 7, 3144-57.
- Tanikawa, C. et al. (2012) Nat Commun 3, 676.
- Lu, M. et al. (2020) J Med Virol 92, 1221-1230.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
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