Antibodies, Proteins, Kits and Reagents for Life Science

281 433 ₸ Product size

100 µl 281 433 ₸

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Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18378-45] to APE1 - ChIP Grade
Suitable for: IHC-P, ICC/IF, Flow Cyt, WB, ChIP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-APE1 antibody [EPR18378-45] - ChIP Grade
See all APE1 primary antibodies
Description

Rabbit monoclonal [EPR18378-45] to APE1 - ChIP Grade
Host species

Rabbit

Tested Applications & Species

Application	Species
ChIP	Human
Flow Cyt	Mouse
ICC/IF	Mouse Human
IHC-P	Mouse Rat Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal brain, fetal heat, fetal kidney and fetal spleen tissue lysate; mouse brain, heart and liver tissue lysate; rat brain, liver and spleen tissue lysate; HCT 116, HeLa, NIH/3T3, HEK293, HepG2, wild-type HAP1, and C6 whole cell lysates. IHC-P: Human ovarian carcinoma tissue; mouse liver tissue; rat liver tissue. ICC/IF: HCT 116 and NIH/3T3 cells. Flow cytometry: NIH/3T3 cells. ChIP: HCT 116 cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18378-45
Isotype

IgG
Research areas

Western blot - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

All lanes : Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474) at 1/1000 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: 35 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 - ab189474 observed at 37 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab189474 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 ab189474 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 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.
Western blot - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

All lanes : Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/2000 dilution

Developed using the ECL technique.

Predicted band size: 35 kDa
Observed band size: 35 kDa

Blocking and dilution buffer: 5% NFDM/TBST

Exposure times:

Lanes 1-3: 1 second

Lane 4: 4 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling APE1 with ab189474 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on tumor cells of human ovarian carcinoma (PMID: 20087352) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling APE1 with ab189474 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling APE1 with ab189474 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on tumor cells of human ovarian carcinoma (PMID: 20087352) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling APE1 with ab189474 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Mainly nuclear staining on hepatocytes of rat liver (PMID: 10643898) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma cell line) cells labeling APE1 with ab189474 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HCT 116 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution.

-ve control : PBS, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution
Immunocytochemistry/ Immunofluorescence - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling APE1 with ab189474 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution.

-ve control : PBS, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution
Western blot - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

All lanes : Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474) at 1/1000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse liver lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat liver lysate
Lane 6 : Rat spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 35 kDa
Observed band size: 35 kDa

Blocking and dilution buffer: 5% NFDM/TBST

Exposure times:

Lanes 1-5: 8 seconds

Lane 6: 4 seconds
Western blot - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

All lanes : Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474) at 1/5000 dilution

Lane 1 : HCT 116 (human colorectal carcinoma cell line) whole cell lysate
Lane 2 : HeLa(human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 4 : C6 (rat brain glioma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 35 kDa
Observed band size: 35 kDa

Blocking and dilution buffer: 5% NFDM/TBST

Exposure times:

Lanes 1 & 2: 1 second

Lane 3: 5 seconds

Lane 4: 3 seconds
Flow Cytometry - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

Flow cytometric analysis of 4% pasraformaldehyde-fixed, 90% methanol-permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cell line labeling APE1 with ab189474 at 1/500 dilution (red) compared with a Isotype control details (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.
ChIP - Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

Chromatin was prepared from HCT 116 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab189474 (blue), and 20µl of Protein A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 5μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR green approach).

ChIP was performed according to the literature (PMID: 23874636).
Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474)

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