APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 (APEX1) antibody

Details for Product No. ABIN4889971, Supplier: Log in to see
Antigen
  • apex1
  • MGC76280
  • MGC79645
  • MGC78928
  • APE
  • APE1
  • APEN
  • APEX
  • APX
  • HAP1
  • REF1
  • Apex
  • REF-1
  • BAP1
  • apex1a
  • hm:zehn1532
  • wu:fc13d11
  • zgc:66204
  • Ref-1
  • apurinic/apyrimidinic endodeoxyribonuclease 1
  • APEX nuclease (multifunctional DNA repair enzyme) 1
  • APEX nuclease (multifunctional DNA repair enzyme) 1 L homeolog
  • DNA-(apurinic or apyrimidinic site) lyase
  • apurinic/apyrimidinic endonuclease 1
  • APEX1
  • apex1
  • apex1.L
  • CNG03640
  • CNH01280
  • nth
  • Spirs_4119
  • Palpr_0103
  • Apex1
Alternatives
anti-Human APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 antibody for Chromatin Immunoprecipitation
Reactivity
Human, Mouse (Murine), Primate, Rat (Rattus)
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84
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Host
Mouse
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Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
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1
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Application
Chromatin Immunoprecipitation (ChIP), Gel Shift (GS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Proximity Ligation Assay (PLA), Simple Western (SimWes), Western Blotting (WB)
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13
9
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1
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1
Options
Supplier
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Supplier Product No.
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Immunogen Purified human APE1 [UniProt# P27695]
Clone 13B8E5C2
Isotype IgG2b
Purification Protein G purified
Antigen
Alternative Name APEX1 / REF-1 (APEX1 Antibody Abstract)
Background Gene Symbol: APEX1
Molecular Weight Theoretical MW: 37 kDa
Gene ID 328
UniProt P27695
Pathways DNA Damage Repair, Chromatin Binding, Cell RedoxHomeostasis, Smooth Muscle Cell Migration, Positive Regulation of Response to DNA Damage Stimulus
Application Notes Western Blot 1:100-1:2000, Simple Western 1:25, Chromatin Immunoprecipitation 1:10-1:500, Immunohistochemistry 1:100, Immunocytochemistry/Immunofluorescence 1:50-1:200, Immunoprecipitation 1:10-1:500, Immunohistochemistry-Paraffin 1:100, Immunohistochemistry-Frozen 1:10-1:500, Gel Super Shift Assays, Proximity Ligation AssayThis APE1 (13B8E5C2) antibody is useful for Western Blot, Chromatin Immunoprecipitation, Immunocytochemistry/Immunofluorescence, Immunoprecipitation and Immunohistochemistry. Gel Super Shift Assays was reported in scientific literature. In WB this antibody detects a single band at 37 kDa. In IHC, it can be competitively inhibited from recognizing the APE1 antigen in tissues using APE1 protein. It can also be used on frozen and fixed-paraffin sections and cytospin preps. In IHC-P, staining was observed in the nucleus of a human breast cancer xenograft. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer ( pH 6.0) is recommended. In Simple Western only 10 - 15 μL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Use in Proximity Ligation Assay reported in scientific literature (PMID 27808278).
Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol Protocol specific for APE1 Antibody Western Blot
1. Gels, Whatman paper, and membranes are soaked in electroblotting buffer (25 mM Tris-HCl, 193 mM glycine, 20 % methanol) for 15 minutes prior to transferring
. Proteins separated on SDS-polyacrylamide gels are transferred onto 0.22 micron nitrocellulose sheets by electroblotting in a Transblot BioRad transfer apparatus in 25 mM Tris, 192 mM Glycine, 20 % Methanol at 150 mA (70 V). The transfer is carried out for 1 hour at 4 degrees C.
. Following protein transfer, the filter is blocked with Blotto [1X TBST (10X TBST = 1.5 M NaCl, 100 mM Tris-HCl, pH 8.0, 0.5 % Tween 20, 2 % NP-40, 0.2 % SDS), 5 % Carnation dried milk, 0.02 % sodium azide] for 1 hour at room temperature on a rotator.
. Dilute NB 100-116 (anti-APE/ref-1) in Blotto and incubate with the filter, at 4 degrees C overnight, on a rotator.
. Wash filter 3 times in 1X TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1 % Tween 20, pH 7.
5) for 10 minutes at 4 degrees C. Secondary antibody (peroxidase conjugated anti-mouse) is incubated with the blot for 30 minutes at room temperature. Cross-reacting proteins are detected using the Chemiluminescence Western Blotting Kit from Boehringer-Mannheim. NOTE: HeLa whole cell extracts were used as a positive control for this antibody. Immunohistochemistry/Immunocytochemistry The description that follows is for cultured cells but can be used for cytospins.
. Split cells into 3.5 cm culture dishes for growth.
. Wash cells with 5 mL PBS.
. Fix cells with approx. 3 mL Histochoice (Amresco) for 30 min (Cryostat tissues for 45 min) or use 10 % formalin for 30 minutes.
. Rinse cells with 5 mL TBS, wipe plate dry leaving a small circle of buffer and cells. Mark with red pencil.
. Pre-block the cells for 30 min. with 10 % goat serum in TBS (200 µL).
. Aspirate blocking solution and add NB 100-116 (anti-APE/ref-1), dilution in 10 % goat serum.
. Incubate in humidified chamber for 3 hours (overnight for tissue at 4 degrees C).
. Incubate the cells with 1:100 diluted secondary antibody (anti-mouse IgG made in goat) in 10 % goat serum and TBS for 1 hour in humidified chamber.
. Wash 2 times with 5 mL TBS for 5 min each.
. Block with ABC solution for 30 min.
. Wash 2 times with 5 mL TBS for 5 min each.
. Incubate with DAB solution until signal develops. Place into dH2O. Add coverslip with Aqua-mount. TBS: 50 mM Tris, 150 mM NaCl, pH 7.5, ABC and DAB solutions: Vector laboratories
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer Tris-Glycine and 0.15M NaCl
Buffer contains: 0.05 % Sodium Azide
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C,-20 °C
Storage Comment Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.
Supplier Images
Western Blotting (WB) image for anti-APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 (APEX1) antibody (ABIN4889971) anti-APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 (APEX1) antibody
Immunohistochemistry (IHC) image for anti-APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 (APEX1) antibody (ABIN4889971) Immunocytochemistry: APE1 Antibody (13B8E5C2) [ABIN151042] - Immunocytochemical detec...
Simple Western (SimWes) image for anti-APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 (APEX1) antibody (ABIN4889971) Simple Western: APE Antibody (13B8E5C2) [ABIN151042] - Simple Western lane view shows...
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 (APEX1) antibody (ABIN4889971) Immunohistochemistry-Paraffin: APE Antibody (13B8E5C2) [ABIN151042] - APE1 antibody w...
Immunofluorescence (IF) image for anti-APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 (APEX1) antibody (ABIN4889971) Immunocytochemistry/Immunofluorescence: APE Antibody (13B8E5C2) [ABIN151042] - HeLa c...
Western Blotting (WB) image for anti-APEX Nuclease (Multifunctional DNA Repair Enzyme) 1 (APEX1) antibody (ABIN4889971) Western Blot: APE Antibody (13B8E5C2) - Whole cell protein from human HeLa, A431, mo...
Product cited in: Antoniali, Serra, Lirussi, Tanaka, DAmbrosio, Zhang, Radovic, Dalla, Ciani, Scaloni, Li, Piazza, Tell: "Mammalian APE1 controls miRNA processing and its interactome is linked to cancer RNA metabolism." in: Nature communications, Vol. 8, Issue 1, pp. 797, 2018 (PubMed). Method employed by authors: Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Ahuja, Jodkowska, Teloni, Bizard, Zellweger, Herrador, Ortega, Hickson, Altmeyer, Mendez, Lopes: "A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells." in: Nature communications, Vol. 7, pp. 10660, 2016 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting

Wang, Cai, Bailey, Reza Saadatzadeh, Ding, Tonsing-Carter, Georgiadis, Zachary Gunter, Long, Minto, Gordon, Sen, Cai, Eitel, Waning, Bringman, Wells, Murray, Sarkaria, Gelbert, Jones, Cohen-Gadol et al.: "Combination therapy in a xenograft model of glioblastoma: enhancement of the antitumor activity of temozolomide by an MDM2 antagonist. ..." in: Journal of neurosurgery, pp. 1-14, 2016 (PubMed).

Lee, Lim, Shin, Choi, Joo, Jeon: "Altered Secretory Activity of APE1/Ref-1 D148E Variants Identified in Human Patients With Bladder Cancer." in: International neurourology journal, Vol. 20, Issue Suppl 1, pp. S30-37, 2016 (PubMed). (Sample species: Human). Further details: Western Blotting

Akhter, Takeda, Nara, Araki, Ishii, Asao, Asao: "Apurinic/Apyrimidinic Endonuclease 1/Redox Factor-1 (Ape1/Ref-1) Modulates Antigen Presenting Cell-mediated T Helper Cell Type 1 Responses." in: The Journal of biological chemistry, Vol. 291, Issue 45, pp. 23672-23680, 2016 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting

Moreli, Santos, Lorenzon-Ojea, Corrêa-Silva, Fortunato, Rocha, Rudge, Damasceno, Bevilacqua, Calderon: "Hyperglycemia Differentially Affects Maternal and Fetal DNA Integrity and DNA Damage Response." in: International journal of biological sciences, Vol. 12, Issue 4, pp. 466-77, 2016 (PubMed).

Kelley, Wikel, Guo, Pollok, Bailey, Wireman, Fishel, Vasko: "Identification and Characterization of New Chemical Entities Targeting Apurinic/Apyrimidinic Endonuclease 1 for the Prevention of Chemotherapy-Induced Peripheral Neuropathy." in: The Journal of pharmacology and experimental therapeutics, Vol. 359, Issue 2, pp. 300-309, 2016 (PubMed). Method employed by authors: Chromatin Immunoprecipitation (ChIP)

Bhakat, Sengupta, Adeniyi, Roychoudhury, Nath, Bellot, Feng, Mantha, Sinha, Qiu, Luxon: "Regulation of limited N-terminal proteolysis of APE1 in tumor via acetylation and its role in cell proliferation." in: Oncotarget, Vol. 7, Issue 16, pp. 22590-604, 2016 (PubMed).

Yang, Sengupta, Hegde, Mitra, Jiang, Holey, Sarker, Tsai, Hegde, Mitra: "Regulation of oxidized base damage repair by chromatin assembly factor 1 subunit A." in: Nucleic acids research, 2016 (PubMed).

Szczesny, Marcatti, Zatarain, Druzhyna, Wiktorowicz, Nagy, Hellmich, Szabo: "Inhibition of hydrogen sulfide biosynthesis sensitizes lung adenocarcinoma to chemotherapeutic drugs by inhibiting mitochondrial DNA repair and suppressing cellular bioenergetics." in: Scientific reports, Vol. 6, pp. 36125, 2016 (PubMed). Method employed by authors: Proximity Ligation Assay (PLA) (Sample species: Human).

Sengupta, Mantha, Song, Roychoudhury, Nath, Ray, Bhakat: "Elevated level of acetylation of APE1 in tumor cells modulates DNA damage repair." in: Oncotarget, Vol. 7, Issue 46, pp. 75197-75209, 2016 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Human).

Roychoudhury, Nath, Song, Hegde, Bellot, Mantha, Sengupta, Ray, Natarajan, Bhakat: "Human Apurinic/Apyrimidinic Endonuclease (APE1) Is Acetylated at DNA Damage Sites in Chromatin, and Acetylation Modulates Its DNA Repair Activity." in: Molecular and cellular biology, Vol. 37, Issue 6, 2016 (PubMed). (Sample species: Human).

Nikitaki, Nikolov, Mavragani, Mladenov, Mangelis, Laskaratou, Fragkoulis, Hellweg, Martin, Emfietzoglou, Hatzi, Terzoudi, Iliakis, Georgakilas: "Measurement of complex DNA damage induction and repair in human cellular systems after exposure to ionizing radiations of varying linear energy transfer (LET)." in: Free radical research, Vol. 50, Issue sup1, pp. S64-S78, 2016 (PubMed). Method employed by authors: Immunofluorescence (fixed cells) (IF/ICC) (Sample species: Human).

Yang, Zhang, Zhang, Wan, Zhang, Huang, Huang, Pu, Pei, Wu, Huang, Huang, Li: "KDM1A triggers androgen-induced miRNA transcription via H3K4me2 demethylation and DNA oxidation." in: The Prostate, Vol. 75, Issue 9, pp. 936-46, 2015 (PubMed). (Sample species: Human). Further details: Chromatin Immunoprecipitation

Pojo, Gonçalves, Xavier-Magalhães, Oliveira, Gonçalves, Correia, Rodrigues, Costa, Pinto, Pinto, Lopes, Reis, Rocha, Sousa, Costa: "A transcriptomic signature mediated by HOXA9 promotes human glioblastoma initiation, aggressiveness and resistance to temozolomide." in: Oncotarget, Vol. 6, Issue 10, pp. 7657-74, 2015 (PubMed). (Sample species: Human). Further details: Western Blotting

Aamann, Norregaard, Kristensen, Stevnsner, Frøkiær: "Unilateral ureteral obstruction induces DNA repair by APE1." in: American journal of physiology. Renal physiology, pp. ajprenal.00613.2014, 2015 (PubMed). (Sample species: Rat (Rattus)). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Bonora, Wieckowsk, Chinopoulos, Kepp, Kroemer, Galluzzi, Pinton: "Molecular mechanisms of cell death: central implication of ATP synthase in mitochondrial permeability transition." in: Oncogene, Vol. 34, Issue 12, pp. 1608, 2015 (PubMed). Method employed by authors: Immunohistochemistry (IHC)

Karmahapatra, Saha, Adhikari, Woodrick, Roy: "Redox regulation of apurinic/apyrimidinic endonuclease 1 activity in Long-Evans Cinnamon rats during spontaneous hepatitis." in: Molecular and cellular biochemistry, Vol. 388, Issue 1-2, pp. 185-93, 2014 (PubMed). (Sample species: Rat (Rattus)). Further details: Western Blotting

Xie, Zhang, Li, Du, Zhou, Yang, Zeng, Li, Wang, Wang: "Functional analysis of the involvement of apurinic/apyrimidinic endonuclease 1 in the resistance to melphalan in multiple myeloma." in: BMC cancer, Vol. 14, pp. 11, 2014 (PubMed). (Sample species: Human). Further details: Western Blotting,Immunoprecipitation

Kim, Koedrith, Lee, Kim, Seo: "Base excision DNA repair defect in thioredoxin-1 (Trx1)-deficient cells." in: Mutation research, Vol. 751-752, pp. 1-7, 2014 (PubMed). (Sample species: Human).

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