EM1801-12

APE1 Mouse Monoclonal Antibody [12H2]

cat.: EM1801-12

Product Type:	Mouse monoclonal IgG2b, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	12H2

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 36 kDa
Isotype:	IgG2b
Immunogen:	Recombinant protein within Human APE1 aa 20-318/318.
Positive control:	HeLa cell lysate, LNCaP cell lysate, HEK-293 cell lysate, HL-60 cell lysate, A431 cell lysate, NIH/3T3 cell lysate, mouse brain tissue lysate, C6 cell lysate, rat testis tissue lysate, A549, human kidney tissue, mouse liver tissue, rat colon tissue, HeLa.
Subcellular location:	Nucleus. Endoplasmic reticulum, Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000-1:5,000 1:100 1:1,000 1:1,000
Uniprot #:	SwissProt: P27695 Human \| P28352 Mouse \| P43138 Rat
Alternative names:	AP endonuclease 1 AP endonuclease class I AP lyase APE 1 APE APE-1 APEN APEX 1 APEX APEX nuclease (multifunctional DNA repair enzyme) 1 Apex nuclease 1 APEX nuclease APEX1 APEX1_HUMAN Apurinic endonuclease Apurinic-apyrimidinic endonuclease 1 Apurinic/apyrimidinic (abasic) endonuclease Apurinic/apyrimidinic endonuclease 1 Apurinic/apyrimidinic exonuclease APX BAP1 Deoxyribonuclease (apurinic or apyrimidinic) DNA (apurinic or apyrimidinic site) lyase DNA-(apurinic or apyrimidinic site) lyase, mitochondrial EC 4.2.99.18 HAP 1 HAP1 Human Apurinic endonuclease 1 MGC139790 Multifunctional DNA repair enzyme Redox factor 1 Redox factor-1 REF 1 REF 1 protein REF-1 REF1 REF1 protein

Images

Fig1: Western blot analysis of APE1 on different lysates with Mouse anti-APE1 antibody (EM1801-12) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: LNCaP cell lysate (20 µg/Lane)
Lane 3: HEK-293 cell lysate (20 µg/Lane)
Lane 4: HL-60 cell lysate (20 µg/Lane)
Lane 5: A431 cell lysate (20 µg/Lane)
Lane 6: NIH/3T3 cell lysate (20 µg/Lane)
Lane 7: Mouse brain tissue lysate (40 µg/Lane)
Lane 8: C6 cell lysate (20 µg/Lane)
Lane 9: Rat testis tissue lysate (40 µg/Lane)

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 1 minute;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1801-12) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of A549 cells labeling APE1 with Mouse anti-APE1 antibody (EM1801-12) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-APE1 antibody (EM1801-12) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-APE1 antibody (EM1801-12) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM1801-12) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-APE1 antibody (EM1801-12) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM1801-12) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Mouse anti-APE1 antibody (EM1801-12) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM1801-12) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig6: Flow cytometric analysis of HeLa cells labeling APE1.

Cells were fixed and permeabilized. Then stained with the primary antibody (EM1801-12, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.