Overview

  • Product name

    Anti-APE1 antibody [EPR4022] - BSA and Azide free
    See all APE1 primary antibodies
  • Description

    Rabbit monoclonal [EPR4022] to APE1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human APE1 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: K562, HepG2, Raji , C6, PC-12, NIH3T3 and Raw264.7 whole cell lysates; Rat brain and spleen lysates; Mouse brain, heart, kidney and spleen lysates IHC-P: Human tonsil, human kidney, human liver, mouse liver, and rat spleen tissues.
  • General notes

    Ab214805 is the carrier-free version of ab92744. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab214805 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab214805 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 Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Target

  • 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
    modifications

    Phosphorylated. 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
  • Database links

  • Alternative names

    • AP endonuclease 1 antibody
    • AP endonuclease class I antibody
    • AP lyase antibody
    • APE 1 antibody
    • APE antibody
    • APE-1 antibody
    • APEN antibody
    • APEX 1 antibody
    • APEX antibody
    • APEX nuclease (multifunctional DNA repair enzyme) 1 antibody
    • Apex nuclease 1 antibody
    • APEX nuclease antibody
    • APEX1 antibody
    • APEX1_HUMAN antibody
    • Apurinic endonuclease antibody
    • Apurinic-apyrimidinic endonuclease 1 antibody
    • Apurinic/apyrimidinic (abasic) endonuclease antibody
    • Apurinic/apyrimidinic endonuclease 1 antibody
    • Apurinic/apyrimidinic exonuclease antibody
    • APX antibody
    • BAP1 antibody
    • Deoxyribonuclease (apurinic or apyrimidinic) antibody
    • DNA (apurinic or apyrimidinic site) lyase antibody
    • DNA-(apurinic or apyrimidinic site) lyase, mitochondrial antibody
    • EC 4.2.99.18 antibody
    • HAP 1 antibody
    • HAP1 antibody
    • Human Apurinic endonuclease 1 antibody
    • MGC139790 antibody
    • Multifunctional DNA repair enzyme antibody
    • Redox factor 1 antibody
    • Redox factor-1 antibody
    • REF 1 antibody
    • REF 1 protein antibody
    • REF-1 antibody
    • REF1 antibody
    • REF1 protein antibody
    see all

Images

  • 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.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 kDa



    Blocking/diluting buffer and concentration:
    5% NFDM /TBST 5% NFDM /TBST

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 kDa



    Blocking/diluting buffer and concentration:
    5% NFDM /TBST 5% NFDM /TBST

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92744).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92744).

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • 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 kDa



    Blocking/diluting buffer and concentration:
    5% NFDM /TBST 5% NFDM /TBST

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92744).

References

This product has been referenced in:

  • 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 . Read more (PubMed: 22497302) »
See 1 Publication for this product

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