Anti-APE1 antibody [EPR4022]

• IHC-P

Unknown

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.

• IHC-P

Unknown

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.

• IHC-P

Unknown

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.

• IHC-P

Unknown

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.

• WB

Lab

Western blot - Anti-APE1 antibody [EPR4022] (AB92744)

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.

All lanes:

Western blot - Anti-APE1 antibody [EPR4022] (ab92744) at 1/10000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

APEX1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HepG2 whole cell lysate at 20 µg

Lane 4:

HEK293 whole cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 37 kDa

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• WB

Lab

Western blot - Anti-APE1 antibody [EPR4022] (AB92744)

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

All lanes:

Western blot - Anti-APE1 antibody [EPR4022] (ab92744) at 0.51 µg/mL

Lane 1:

Rat brain lysate at 20 µg

Lane 2:

Rat heart lysate at 20 µg

Lane 3:

Rat liver lysate at 20 µg

Lane 4:

Rat spleen lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 0.05 µg/mL

Predicted band size: 35 kDa

Observed band size: 36 kDa

false

• WB

Lab

Western blot - Anti-APE1 antibody [EPR4022] (AB92744)

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

All lanes:

Western blot - Anti-APE1 antibody [EPR4022] (ab92744) at 0.51 µg/mL

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

Mouse heart lysate at 20 µg

Lane 3:

Mouse kidney lysate at 20 µg

Lane 4:

Mouse spleen lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 0.05 µg/mL

Predicted band size: 35 kDa

Observed band size: 36 kDa

false

• WB

Lab

Western blot - Anti-APE1 antibody [EPR4022] (AB92744)

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

All lanes:

Western blot - Anti-APE1 antibody [EPR4022] (ab92744) at 0.51 µg

Lane 1:

C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 0.05 µg/mL

Predicted band size: 35 kDa

false

• WB

Lab

Western blot - Anti-APE1 antibody [EPR4022] (AB92744)

All lanes:

Western blot - Anti-APE1 antibody [EPR4022] (ab92744) at 1/20000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 36 kDa

false