Immunofluorescence staining using MA1-2022, treatment with paraquat and iron induces MnSOD and Phosphorylation of H2AX in RAW 264.7 macrophages. Cells were treated for 20 hours with paraquat (500 µM) and iron (200 µg/ml) and stained with anti-Phospho-H2AX antibody.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Bovine, Human, Not Applicable|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Synthetic peptide sequence surrounding phosphorylated Ser140|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Flow Cytometry (Flow)||1µg per 10^6 cells|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunofluorescence (IF)||2-4 ug/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:400|
|Western Blot (WB)||1 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-2022 was used in IHC to successfully detect H2A.X pSer140 in postnatal mouse lung section.
MA1-2022 detects human and mouse phosphorylated H2AX.
MA1-2022 has been used successfully in Western blot, immunofluorescence, and ELISA procedures. By Western blot this antibody detects ~17 kDa protein representing phosphorylated H2AX in gamma irradiated HeLa cell lysate. In immunofluorescence procedures, MA1-2022 recognizes phosphorylated H2AX in gamma irradiated HeLa cells. ELISA of phosphorylated H2AX can also be performed using MA1-2022. MA1-2022 was used in IHC to successfully detect H2A.X pSer140 in postnatal mouse lung section.
The MA1-2022 immunogen is a synthetic peptide sequence surrounding phosphorylated Ser140. Some references call Ser140, by the name Ser139.
This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
Histones are basic nuclear proteins that are responsible for the structure of eukaryotic chromosomal fiber. H2AX is a member of the histone H2A family which is one of the four core histones making up the nucleosome core particle. In eukaryotes DNA double strand breaks (DSBs) have been shown to trigger the phosphorylation of serine 139 at the carboxy terminus of histone H2AX resulting in gamma-H2AX. The phosphorylation of H2AX can be detected by Western blotting or immunofluorescence, revealing the frequency of DSBs. The phosphatidylinositol 3-kinases have been implicated in H2AX phosphorylation, but it is unclear if ATM is the primary H2AX kinase or if other members of the family such as DNA-PK and ATR contribute in a similar manner.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Elevated Expression of Hepatoma Up-Regulated Protein Inhibits ¿-Irradiation-Induced Apoptosis of Prostate Cancer Cells.
MA1-2022 was used in immunocytochemistry and western blot to characterize inhibition of gamma-irradiation-induced apoptosis of prostate cancer cells by elevated expression of hepatoma up-regulated protein
|Hassan M,El Khattouti A,Ejaeidi A,Ma T,Day WA,Espinoza I,Vijayakumar S,Gomez CR||Journal of cellular biochemistry (117:1308)||2016|
High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents (HPHC).
MA1-2022 was used in immunocytochemistry to evaluate the toxicological effects of harmful and potentially harmful constituents (HPHC) by utilizing a high content screening analysis
|Marescotti D,Gonzalez Suarez I,Acali S,Johne S,Laurent A,Frentzel S,Hoeng J,Peitsch MC||Journal of visualized experiments : JoVE (null:null)||2016|
A high-throughput chemical screen reveals that harmine-mediated inhibition of DYRK1A increases human pancreatic beta cell replication.
MA1-2022 was used in immunocytochemistry to identify analogs of harmine as a new class of human beta cell mitogenic compounds and identify the pathways they activate.
|Wang P,Alvarez-Perez JC,Felsenfeld DP,Liu H,Sivendran S,Bender A,Kumar A,Sanchez R,Scott DK,Garcia-Ocaña A,Stewart AF||Nature medicine (21:383)||2015|
Comparative analysis of the cytotoxic effects of okadaic acid-group toxins on human intestinal cell lines.
MA1-2022 was used in immunocytochemistry to compared the toxicological effects of okadaic acid and dinophysistoxin 1 and 2
|Ferron PJ,Hogeveen K,Fessard V,Le Hégarat L||Marine drugs (12:4616)||2014|
The NR4A2 nuclear receptor is recruited to novel nuclear foci in response to UV irradiation and participates in nucleotide excision repair.
MA1-2022 was used in immunocytochemistry to study the mechanism underlying the role of the NR4A2 nuclear receptor in nucleotide excision repair in response to damage caused by UV irradiation
|Jagirdar K,Yin K,Harrison M,Lim W,Muscat GE,Sturm RA,Smith AG||PloS one (8:null)||2013|
Cytotoxicity, fractionation and dereplication of extracts of the dinoflagellate Vulcanodinium rugosum, a producer of pinnatoxin G.
MA1-2022 was used in immunocytochemistry to isolate and identify N-carboxy-methyl-smenospongine as the likely cytotoxic and genotoxic component of extracts of Vulcanodium rugosum
|Geiger M,Desanglois G,Hogeveen K,Fessard V,Leprêtre T,Mondeguer F,Guitton Y,Hervé F,Séchet V,Grovel O,Pouchus YF,Hess P||Marine drugs (11:3350)||2013|
Profiling dose-dependent activation of p53-mediated signaling pathways by chemicals with distinct mechanisms of DNA damage.
MA1-2022 was used in western blot to develop high throughput methods to assess proof-of-concept in vitro-only risk assessments
|Clewell RA,Sun B,Adeleye Y,Carmichael P,Efremenko A,McMullen PD,Pendse S,Trask OJ,White A,Andersen ME||Toxicological sciences : an official journal of the Society of Toxicology (142:56)||2014|
Development of a two-step chromatography procedure that allows the purification of a high-purity anti-histone H1 monoclonal immunoglobulin M antibody with immunosuppressant activity.
MA1-2022 was used in ELISA to generate and evaluate a monoclonal antibody with immunosuppressant activity
|Shimada Y,Goto T,Kawamoto S,Kiso T,Katayama A,Yamanaka Y,Aki T,Chiang KC,Nakano T,Goto S,Chen CL,Ohmori N,Ono K,Sato S||Biomedical chromatography : BMC (22:13)||2008|