Immunofluorescent analysis of APE1 (green) showing staining in the in the nucleus of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an APE1 monoclonal antibody (Product # MA1-440) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Dog, Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Full-length recombinant human APE protein.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||Assay Dependent|
|Flow Cytometry (Flow)||2-5 µg/mL|
|Immunohistochemistry (Frozen) (IHC (F))||1/5000|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:1000|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* 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.
|Immunohistochemistry (IHC)||See 1 publications below|
MA1-440 detects apurinic/apyrimidinic endonuclease (APE/ref-1) from human, non human primate, canine, mouse and rat samples.
MA1-440 has been successfully used in Western blot, immunohistochemical, flow cytometry and immunoprecipitation procedures. By Western blot, this antibody detects a 36 kDa protein representing APE/ref-1 in HeLa cell lysate. Immunohistochemical staining of APE/ref-1 in a variety of normal and cancerous human tissues, including ovaries, cervix, prostate, germ cell tumors, osteosarcomas, and rhabdosarcomas, with MA1-440 yields a staining pattern consistent with nuclear staining.
The MA1-440 immunogen is full-length, recombinant human APE protein.
Mammalian apurinic/apyrimidinic endonuclease (APE/ref-1) is a multifunctional, bipartite enzyme that plays an important role in numerous, cellular functions. APE is responsible for repairing abasic sites in DNA and in regulating the redox state of other proteins that play roles in oxidative signaling, transcription factor regulation (Fos, Jun, NF-kB, Myb, HIF-1 alpha, CREB, Pax), cell cycle control (p53), and apoptosis. The most common form of DNA damage is the creation of abasic sites which are brought about through spontaneous loss or oxidative DNA damage, through chemically initiated hydrolysis (chemotherapy), ionizing radiation, UV irradiation, oxidizing agents, and removal of modified bases by DNA glycosylases.
APE is differentially expressed during development and in different tissues. This protein has diverse subcellular localization patterns which support the possibility of its interaction with numerous, other cellular proteins in addition to DNA repair within the nucleus. Regulation of APE by phosphorylation is mediated, at least in part, by casein kinase II. Increases in APE message and protein levels are observed upon the reintroduction of oxygen to hypoxic cells, and in some malignant tissue relative to normal tissue. Decreases in APE expression have been associated with the induction of cellular apoptosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Studies of apurinic/apyrimidinic endonuclease/ref-1 expression in epithelial ovarian cancer: correlations with tumor progression and platinum resistance.
MA1-440 was used in immunohistochemistry to investigate the correlations of apurinic/apyrimidinic endonuclease/ref-1 expression with tumor progression and platinum resistance in epithelial ovarian cancer
|Freitas S,Moore DH,Michael H,Kelley MR||Clinical cancer research : an official journal of the American Association for Cancer Research (9:4689)||2003|