Recombinant Anti-APE1 antibody [EPR4022] (ab92744)
![Western blot - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-380868-anti-ape1-antibody-epr4022-western-blot-hap1-wt-hap1-apex1-knockout-hepg2-hek293.jpg "Western blot - Anti-APE1 antibody [EPR4022] (ab92744)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4022] to APE1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-APE1 antibody [EPR4022]
See all APE1 primary antibodies -
Description
Rabbit monoclonal [EPR4022] to APE1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human APE1 aa 1-100 (N terminal). The exact sequence is proprietary.
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Positive control
- WB: Wild-type HAP1, K562, HepG2, HEK293, Raji , C6, PC-12, NIH3T3 and Raw264.7 whole cell lysates; rat brain and spleen lysates; mouse brain, heart, kidney and spleen tissue lysates. IHC-P: Human tonsil, human kidney, human liver, mouse liver, and rat spleen tissues.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4022 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92744 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/10000 - 1/50000. Predicted molecular weight: 36 kDa.
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| IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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| Notes |
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WB
1/10000 - 1/50000. Predicted molecular weight: 36 kDa. |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
Target
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Function
Multifunctional protein that plays a central role in the cellular response to oxidative stress. The two major activities of APEX1 in 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. -
Sequence similarities
Belongs to the DNA repair enzymes AP/ExoA family. -
Domain
The N-terminus contains the redox activity while the C-terminus exerts the DNA AP-endodeoxyribonuclease activity; both function are independent in their actions. An unconventional mitochondrial targeting sequence (MTS) is harbored within the C-terminus, that appears to be masked by the N-terminal sequence containing the nuclear localization signal (NLS), that probably blocks the interaction between the MTS and Tom proteins. -
Post-translational
modificationsPhosphorylated. Phosphorylation by kinase PKC or casein kinase CK2 results in enhanced redox activity that stimulates binding of the FOS/JUN AP-1 complex to its cognate binding site. AP-endodeoxyribonuclease activity is not affected by CK2-mediated phosphorylation. Phosphorylation of Thr-233 by CDK5 reduces AP-endodeoxyribonuclease activity resulting in accumulation of DNA damage and contributing to neuronal death.
Acetylated on Lys-6 and Lys-7. Acetylation is increased by the transcriptional coactivator EP300 acetyltransferase, genotoxic agents like H(2)O(2) and methyl methanesulfonate (MMS). Acetylation increases its binding affinity to the negative calcium response element (nCaRE) DNA promoter. The acetylated form induces a stronger binding of YBX1 to the Y-box sequence in the MDR1 promoter than the unacetylated form. Deacetylated on lysines. Lys-6 and Lys-7 are deacetylated by SIRT1.
Cleaved at Lys-31 by granzyme A to create the mitochondrial form; leading in reduction of binding to DNA, AP endodeoxynuclease activity, redox activation of transcription factors and to enhanced cell death. Cleaved by granzyme K; leading to intracellular ROS accumulation and enhanced cell death after oxidative stress.
Cys-65 and Cys-93 are nitrosylated in response to nitric oxide (NO) and lead to the exposure of the nuclear export signal (NES).
Ubiquitinated by MDM2; leading to translocation to the cytoplasm and proteasomal degradation. -
Cellular localization
Mitochondrion. The cleaved APEX2 is only detected in mitochondria (By similarity). Translocation from the cytoplasm to the mitochondria is mediated by ROS signaling and cleavage mediated by granzyme A. Tom20-dependent translocated mitochondrial APEX1 level is significantly increased after genotoxic stress and Nucleus. Nucleus, nucleolus. Nucleus speckle. Endoplasmic reticulum. Cytoplasm. Detected in the cytoplasm of B-cells stimulated to switch (By similarity). Colocalized with SIRT1 in the nucleus. Colocalized with YBX1 in nuclear speckles after genotoxic stress. Together with OGG1 is recruited to nuclear speckles in UVA-irradiated cells. Colocalized with nucleolin and NPM1 in the nucleolus. Its nucleolar localization is cell cycle dependent and requires active rRNA transcription. Colocalized with calreticulin in the endoplasmic reticulum. Translocation from the nucleus to the cytoplasm is stimulated in presence of nitric oxide (NO) and function in a CRM1-dependent manner, possibly as a consequence of demasking a nuclear export signal (amino acid position 64-80). S-nitrosylation at Cys-93 and Cys-310 regulates its nuclear-cytosolic shuttling. Ubiquitinated form is localized predominantly in the cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 328 Human
- Entrez Gene: 11792 Mouse
- Entrez Gene: 79116 Rat
- Omim: 107748 Human
- SwissProt: P27695 Human
- SwissProt: P28352 Mouse
- SwissProt: P43138 Rat
- Unigene: 73722 Human
see all -
Alternative names
- AP endonuclease 1 antibody
- AP endonuclease class I antibody
- AP lyase antibody
see all
Images
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Western blot - Anti-APE1 antibody [EPR4022] (ab92744)All lanes : Anti-APE1 antibody [EPR4022] (ab92744) at 1/10000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : APEX1 knockout HAP1 whole cell lysate
Lane 3 : HepG2 whole cell lysate
Lane 4 : HEK293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab92744 observed at 37 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab92744 was shown to react with APEX1 in HAP1 wild-type cells in Western blot. Loss of signal was observed when APEX1 knockout sample was used. HAP1 wild-type and APEX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab92744 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-339331-ab92744WB1.jpg)
Western blot - Anti-APE1 antibody [EPR4022] (ab92744)All lanes : Anti-APE1 antibody [EPR4022] (ab92744) at 0.51 µg/ml (purified)
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/diluting buffer and concentration:
5% NFDM /TBST 5% NFDM /TBST -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-8-anti-ape1-antibody-epr4022-immunohistochemistry-spleen-rat.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR4022] (ab92744)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue sections labeling APE1 with purified ab92744 at 1/4000 dilution (0.12 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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![Western blot - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-365472-anti-ape1-antibody-epr4022-westernblot-human.jpg)
Western blot - Anti-APE1 antibody [EPR4022] (ab92744)All lanes : Anti-APE1 antibody [EPR4022] (ab92744) at 1/20000 dilution (Purified)
Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa -
![Western blot - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-339333-ab92744WB3.jpg)
Western blot - Anti-APE1 antibody [EPR4022] (ab92744)All lanes : Anti-APE1 antibody [EPR4022] (ab92744) at 0.51 µg (purified)
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 2 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml
Predicted band size: 36 kDaBlocking/diluting buffer and concentration:
5% NFDM /TBST 5% NFDM /TBST -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-7-anti-ape1-antibody-epr4022-immunohistochemistry-liver-mouse.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR4022] (ab92744)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling APE1 with purified ab92744 at 1/4000 dilution (0.12 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-6-anti-ape1-antibody-epr4022-immunohistochemistry-liver-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR4022] (ab92744)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling APE1 with purified ab92744 at 1/4000 dilution (0.12 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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![Western blot - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-339332-ab92744WB2.jpg)
Western blot - Anti-APE1 antibody [EPR4022] (ab92744)All lanes : Anti-APE1 antibody [EPR4022] (ab92744) at 0.51 µg/ml (purified)
Lane 1 : Rat brain lysate
Lane 2 : Rat heart lysate
Lane 3 : Rat liver lysate
Lane 4 : Rat spleen lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/diluting buffer and concentration:
5% NFDM /TBST 5% NFDM /TBST -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-107970-anti-ape1-antibody-epr4022-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR4022] (ab92744)Unpurified ab92744, at a 1/100 dilution, staining APE1 in formalin fixed, paraffin embedded (1) Human tonsil tissue and (2)Human kidney tissue by Immunohistochemistry. Detection: DAB staining.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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![Anti-APE1 antibody [EPR4022] (ab92744)](https://www.abcam.com/ps/products/92/ab92744/Images/ab92744-11-benefits-of-recombinant-antibodies.png)
Anti-APE1 antibody [EPR4022] (ab92744)
Protocols
Datasheets and documents
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Datasheet download
References (7)
ab92744 has been referenced in 7 publications.
- Dang L et al. Comparison of gene disruption induced by cytosine base editing-mediated iSTOP with CRISPR/Cas9-mediated frameshift. Cell Prolif 53:e12820 (2020). PubMed: 32350961
- Kharat SS et al. Degradation of 5hmC-marked stalled replication forks by APE1 causes genomic instability. Sci Signal 13:N/A (2020). PubMed: 32817374
- Bottardi S et al. MNDA controls the expression of MCL-1 and BCL-2 in chronic lymphocytic leukemia cells. Exp Hematol 88:68-82.e5 (2020). PubMed: 32682001
- Jiang ST et al. Bu-Fei decoction and modified Bu-Fei decoction inhibit the growth of non-small cell lung cancer, possibly via inhibition of apurinic/apyrimidinic endonuclease 1. Int J Mol Med 41:2128-2138 (2018). PubMed: 29393411
- Kovalchuk A et al. Chemo brain or tumor brain - that is the question: the presence of extracranial tumors profoundly affects molecular processes in the prefrontal cortex of TumorGraft mice. Aging (Albany NY) 9:1660-1676 (2017). WB ; Mouse . PubMed: 28758896
- Nassour H et al. Peroxiredoxin 1 interacts with and blocks the redox factor APE1 from activating interleukin-8 expression. Sci Rep 6:29389 (2016). Human . PubMed: 27388124
- Goula AV et al. The nucleotide sequence, DNA damage location, and protein stoichiometry influence the base excision repair outcome at CAG/CTG repeats. Biochemistry 51:3919-32 (2012). WB . PubMed: 22497302
