Western Blot (WB)
(Figure 1. Western blot analysis of APE1 using anti-APE1 antibody (M00627). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysates, Lane 2: human MCF-7 whole cell lysates,Lane 3: human COLO-320 whole cell lysates, Lane 4: human HepG2 whole cell lysates, Lane 5: Rabbit IgG,Lane 6: Marker 1113,Lane 7: human A549 whole cell lysates,Lane 8: human PANC-1 whole cell lysates,Lane 9: human 22RV1 whole cell lysates,Lane 10: human MDA-MB-453 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-APE1 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a Biotin Conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system.)
Mouse anti-Human APE1 Monoclonal Antibody | anti-APE1 antibody
Anti-APE1 Antibody (monoclonal, 5C11)
IHC-P: 0.5-1 mug/ml
IHC-F: 0.5-1 mug/ml
ICC: 0.5-1 mug/ml
FC/FACS: 1-3ug/1x106 cells
Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections.
Avoid repeated freezing and thawing.
Western Blot (WB)
(Figure 1. Western blot analysis of APE1 using anti-APE1 antibody (M00627). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysates, Lane 2: human MCF-7 whole cell lysates,Lane 3: human COLO-320 whole cell lysates, Lane 4: human HepG2 whole cell lysates, Lane 5: Rabbit IgG,Lane 6: Marker 1113,Lane 7: human A549 whole cell lysates,Lane 8: human PANC-1 whole cell lysates,Lane 9: human 22RV1 whole cell lysates,Lane 10: human MDA-MB-453 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-APE1 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a Biotin Conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system.)
Western Blot (WB)
(Figure 1. Western blot analysis of APE1 using anti-APE1 antibody (M00627). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysates, Lane 2: human MCF-7 whole cell lysates,Lane 3: human COLO-320 whole cell lysates, Lane 4: human HepG2 whole cell lysates, Lane 5: Rabbit IgG,Lane 6: Marker 1113,Lane 7: human A549 whole cell lysates,Lane 8: human PANC-1 whole cell lysates,Lane 9: human 22RV1 whole cell lysates,Lane 10: human MDA-MB-453 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-APE1 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a Biotin Conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system.)
Background: APEX1, also called apurinic endonuclease (APE), is a DNA repair enzyme having apurinic/apyrimidinic (AP) endonuclease, 3-prime, 5-prime-exonuclease, DNA 3-prime repair diesterase, and DNA 3-prime-phosphatase activities. The human APEX1 gene consists of 5 exons spanning 2.64 kb and exists as a single copy in the haploid genome. Using in situ hybridization, the APEX1 gene is mapped to 14q11.2-q12. The predicted APEX1 protein, which contained probable nuclear transport signals, was identified as a member of a family of DNA repair enzymes found in lower organisms. The abundance of the large form of APEX1 was increased in leiomyoma extracts relative to myometrial tissue extracts, and the large form was dominant in cell lines derived from leiomyosarcomas. The exonuclease activity of nuclear APEX1 can remove the anti-HIV nucleoside analogs AZT and D4T from the 3-prime terminus of a nick more efficiently than can cytosolic exonucleases.
NCBI and Uniprot Product Information
NCBI Description
Apurinic/apyrimidinic (AP) sites occur frequently in DNA molecules by spontaneous hydrolysis, by DNA damaging agents or by DNA glycosylases that remove specific abnormal bases. AP sites are pre-mutagenic lesions that can prevent normal DNA replication so the cell contains systems to identify and repair such sites. Class II AP endonucleases cleave the phosphodiester backbone 5' to the AP site. This gene encodes the major AP endonuclease in human cells. Splice variants have been found for this gene; all encode the same protein. [provided by RefSeq, Jul 2008]
Uniprot Description
Multifunctional protein that plays a central role in the cellular response to oxidative stress. The two major activities of APEX1 are DNA repair and redox regulation of transcriptional factors. Functions as a apurinic/apyrimidinic (AP) endodeoxyribonuclease in the DNA base excision repair (BER) pathway of DNA lesions induced by oxidative and alkylating agents. Initiates repair of AP sites in DNA by catalyzing hydrolytic incision of the phosphodiester backbone immediately adjacent to the damage, generating a single-strand break with 5'-deoxyribose phosphate and 3'-hydroxyl ends. Does also incise at AP sites in the DNA strand of DNA/RNA hybrids, single-stranded DNA regions of R-loop structures, and single-stranded RNA molecules. Has a 3'-5' exoribonuclease activity on mismatched deoxyribonucleotides at the 3' termini of nicked or gapped DNA molecules during short-patch BER. Possesses a DNA 3' phosphodiesterase activity capable of removing lesions (such as phosphoglycolate) blocking the 3' side of DNA strand breaks. May also play a role in the epigenetic regulation of gene expression by participating in DNA demethylation. Acts as a loading factor for POLB onto non-incised AP sites in DNA and stimulates the 5'-terminal deoxyribose 5'-phosphate (dRp) excision activity of POLB. Plays a role in the protection from granzymes-mediated cellular repair leading to cell death. Also involved in the DNA cleavage step of class switch recombination (CSR). On the other hand, APEX1 also exerts reversible nuclear redox activity to regulate DNA binding affinity and transcriptional activity of transcriptional factors by controlling the redox status of their DNA-binding domain, such as the FOS/JUN AP-1 complex after exposure to IR. Involved in calcium-dependent down-regulation of parathyroid hormone (PTH) expression by binding to negative calcium response elements (nCaREs). Together with HNRNPL or the dimer XRCC5/XRCC6, associates with nCaRE, acting as an activator of transcriptional repression. Stimulates the YBX1-mediated MDR1 promoter activity, when acetylated at Lys-6 and Lys-7, leading to drug resistance. Acts also as an endoribonuclease involved in the control of single-stranded RNA metabolism. Plays a role in regulating MYC mRNA turnover by preferentially cleaving in between UA and CA dinucleotides of the MYC coding region determinant (CRD). In association with NMD1, plays a role in the rRNA quality control process during cell cycle progression. Associates, together with YBX1, on the MDR1 promoter. Together with NPM1, associates with rRNA. Binds DNA and RNA.
Research Articles on APE1
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Product Notes
The APE1 apex1 (Catalog #AAA1751998) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-APE1 Antibody (monoclonal, 5C11) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's APE1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC) Frozen/Paraffin, Immunocytochemistry (ICC), Flow Cytometry (FC/FACS). WB: 0.1-0.5 mug/ml IHC-P: 0.5-1 mug/ml IHC-F: 0.5-1 mug/ml ICC: 0.5-1 mug/ml FC/FACS: 1-3ug/1x106 cells Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. Researchers should empirically determine the suitability of the APE1 apex1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "APE1, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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