Antibodies, Proteins, Kits and Reagents for Life Science

355 142 ₸ Product size

100 µl 355 142 ₸

Order now and get it on Tuesday March 02, 2021

Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR17929] to Oct4 - ChIP Grade
Suitable for: WB, ICC/IF, IP, IHC-P, ChIP, Flow Cyt
Reacts with: Mouse, Rat, Human

Product name

Anti-Oct4 antibody [EPR17929] - ChIP Grade
See all Oct4 primary antibodies
Description

Rabbit monoclonal [EPR17929] to Oct4 - ChIP Grade
Host species

Rabbit
Specificity

Oct4 is highly expressed in undifferentiated embryonic stem cells and cancer stem cell-like cells (PMID: 26013162, 21826175). ab181557 can’t detect the target band in undifferentiated cancer cell lines with low expression level of Oct4, such as HeLa, HEK-293, MDA-MB-231, HepG2, Huh7, HCT-116 and PANC-1 (PMID: 21975933, 29789579, 25625591, 26059097, 23928699, 27344963, 25837691, 29254202, 28854261, 27996162), even at the dilution of 1:200.

Tested Applications & Species

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

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: NCCIT, F9, and NTERA-2 cl.D1 whole cell lysates. IHC-P: Human seminoma and dysgerminoma of ovary tissues. ICC/IF: NCCIT cells. IP: NCCIT whole cell extract. ChIP: Chromatin prepared from F9 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.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17929
Isotype

IgG
Research areas

ChIP - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Chromatin was prepared from F9 (Mouse embyro testicular cancer cell line) 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 ab181557 (red, and 20µl of Anti rabbit IgG sepharose beads. 5μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

“pro” stands for promoter region, while “NC2” stands for negative control which is negative loci at the promoter region.
Western blot - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

All lanes : Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557) at 1/2000 dilution

Lane 1 : NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 5 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 6 : Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 7 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates
Lane 8 : PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 39 kDa
Observed band size: 45 kDa
why is the actual band size different from the predicted?

Exposure time: 3 seconds

Blocking and diluting buffer: 5% NFDM/TBST

Oct4 is highly expressed in undifferentiated embryonic stem cells and cancer stem cell-like cells (PMID: 26013162, 21826175). ab181557 can’t detect the target band in undifferentiated cancer cell lines with low expression level of Oct4, such as HeLa, HEK-293, MDA-MB-231, HepG2, Huh7, HCT-116 and PANC-1 (PMID: 21975933, 29789579, 25625591, 26059097, 23928699, 27344963, 25837691, 29254202, 28854261, 27996162), even at the dilution of 1:200.
Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (Human pluripotent embryonic carcinoma) cells (positive cell line) or NIH/3T3 (Mouse embyro fibroblast) cells (negative cell line) labeling Oct4 with ab181557 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on NCCIT cell line. Negative expression in NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab181557 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557) This image is courtesy of an Abreview submitted by Joanne Lacey.

ab181557 staining Oct4 in Human embryonic stem cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with formaldehyde , permeabilized with 0.1% Triton in PBS for 1 hour and blocked with 10% Serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/200 in PBS with 0.1% Tween20) for 16 hours at 4°C. A monoclonal Goat Anti-rabbit Alexa Fluor^® 594 was used as the secondary antibody at 1/200 dilution.
See Abreview
Immunoprecipitation - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Oct4 was immunoprecipitated from 1mg of NCCIT (Human pluripotent embryonic carcinoma) whole cell extract with ab181557 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab181557 at 1/10000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: NCCIT whole cell extract 10 µg (Input). Lane 2: ab181557 IP in NCCIT whole cell extract. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181557 in NCCIT whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
Western blot - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

All lanes : Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557) at 1/1000 dilution

Lane 1 : NCCIT (Human pluripotent embryonic carcinoma) whole cell lysate
Lane 2 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
Lane 3 : NTERA-2 cl.D1 (Human malignant pluripotent embryonic carcinoma) whole cell lysate
Lane 4 : Mouse testis lysate
Lane 5 : Human hippocampus lysate
Lane 6 : Human cerebellum lysate
Lane 7 : Human testis lysate
Lane 8 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 39 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Low levels of expression in adult tissues
Flow Cytometry - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

ab181557 staining OCT-4 in the human cell line NCCIT (human pluripotent embryonal carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/70. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody (Red).

Isotype control: Rabbit IgG monoclonal [EPR25A] ab172730 (Black).

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Immunohistochemical analysis of paraffin-embedded Human dysgerminoma of ovary tissue labeling Oct4 with ab181557 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weak cytoplasmic staining on cancer cells of Human dysgerminoma of ovary is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Oct4 with ab181557 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Negative staining on Human breast cancer. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Oct4 with ab181557 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Negative staining on adult Human testis. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Oct4 with ab181557 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Negative staining on adult mouse kidney. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Oct4 with ab181557 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Negative staining on adult rat testis. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Anti-Oct4 antibody [EPR17929] - ChIP Grade (ab181557)

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