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Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)
Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)
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Western blot
Western blot
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Immunocytochemistry/ Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)
Immunocytochemistry/ Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)
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Product Name 

Oct4 antibody - ChIP Grade

 

See all Oct4 antibodies (14)...

Product type 

Primary antibodies

Description 

Rabbit polyclonal to Oct4 - ChIP Grade

Immunogen 

Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human Oct4.

(Peptide available as ab20650.)

Reacts with 
(species key)

Hu, Ms

Specificity 

IHC: This antibody is not suitable for staining of mouse paraformaldehyde-fixed frozen sections. We have no available data on the performance of the antibody using alternative conditions or target species in IHC.

Recognises a band of the expected size in mouse and human. Recognises an additional band in mouse but not in human.

Tested applications 
(see key)

ChIP, Flow Cyt, ICC/IF, IHC-P, WB


Abreviews 

Customer reviews
(see key)


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• Flow Cyt

1 Abreview

- average rating: 5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)

• ICC/IF

1 Abreview

- average rating: 5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)

• IHC-P

2 Abreviews

- average rating: 5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)5 stars (out of 5)


IHC-P
(read Abreview)


Flow Cyt
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ICC/IF
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Application notes 
(see key)

Recommended dilutions
ChIP: Use at an assay dependent dilution. See reference PMID 18490265.
Flow Cyt: 1/100 (see Abreview)
ICC/IF: Use at a concentration of 1 - 5 µg/ml.
IHC-P: 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 39 kDa). Can be blocked with Oct4 peptide (ab20650).

Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

Positive control 
(see definition)

Human ES cell lysate

Cellular localization 

Nuclear

Research areas 

Developmental Biology >> Embryogenesis >> Embryonic stem cells >> Surface molecules
Developmental Biology >> Reproduction >> Germ cell markers
Chromatin and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Stem Cells >> Germline Stem Cells >> Embryonic Germ Cells
Stem Cells >> Embryonic Stem Cells >> Intracellular
Chromatin and Nuclear Signaling >> Transcription >> Domain Families >> Developmental Families >> OCT
Neuroscience >> Cell Type Marker >> Glia marker >> Oligodendrocyte marker

Relevance 

Members of Oct family of transcription factors specifically interact with Octamer motif ATGCAAAT, a regulatory element important for tissue- and cell-specific transcription as well as for transcription of a number of housekeeping genes. All of the members of the Oct family contain two highly conserved domains which are separated by 14-26 variable amino acids. These include the POU homeodomain and the POU-specific domain. Both are required for DNA binding and are involved in protein-protein interactions. Evidences indicate that regulation of Oct family transcription factors occurs at the level of phosphorylation. Oct4 is renowned as a transcription factor expressed by undifferentiated embryonic stem cells and embryonic germ cells.

 

Alternative names Oct4 antibody - ChIP Grade (ab19857)

Database links 

The links below go to external sites and will open in a new browser window

Entrez Gene

    

5460    (Human)
18999    (Mouse)
294562    (Rat)

GeneCard

    

GC06M031238    (Human)

Omim

    

164177    (Human)

SwissProt

    

Q01860    (Human)
P20263    (Mouse)

Unigene

    

249184    (Human)

Raised in 

Rabbit

Clonality 

Polyclonal

Isotype 

IgG

Purity 

Immunogen affinity purified

Storage buffer 

Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4

Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS

Form 

Liquid

Concentration 

Batch dependent within range: 0.10 - 0.90 mg/ml

Concentration of lot no.     is  

Concentration not available for this lot.

Find concentration of your lot :

Storage instructions 

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

 

At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Oct4 antibody - ChIP Grade is on this datasheet. But please do contact us if you would like any reassurance!


See below for Oct4 antibody - ChIP Grade images, references, products related to ab19857 and other tools.

 

Oct4 antibody - ChIP Grade images:

Product image  Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)

Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)

The image shows a colony of differentiating Human Embryonic Stem Cells double-stained with anti-Oct4 antibody ab19857 (green) and Sox17 antibody (red). The nuclei of Oct4-positive undifferentiated hESCs stained bright green. Staining was restricted to the nuclei. Oct4-positive nuclei were Sox17-negative, and vice versa. This antibody can be used as a marker of Oct4-positive undifferentiated Human Embryonic Stem Cells.

Ludovic Vallier, University of Cambridge

 

Product image  Western blot

Western blot

All lanes : Oct4 antibody - ChIP Grade (ab19857) at 1 µg/ml

Lane 1 : MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (ab27198)
Lane 2 : MEL-2 (Human embryonic stem cell, female cell line) Whole Cell Lysate (ab27196)
Lane 3 : IOUD2 (Mouse embryonic stem cell, selected for Oct4 expression cell line) Whole Cell Lysate (ab27202)
Lane 4 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate (ab27193)
Lane 5 : MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (ab27198) with Oct4 peptide (ab20650) at 1 µg/ml
Lane 6 : MEL-2 (Human embryonic stem cell, female cell line) Whole Cell Lysate (ab27196) with Oct4 peptide (ab20650) at 1 µg/ml
Lane 7 : IOUD2 (Mouse embryonic stem cell, selected for Oct4 expression cell line) Whole Cell Lysate (ab27202) with Oct4 peptide (ab20650) at 1 µg/ml
Lane 8 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate (ab27193) with Oct4 peptide (ab20650) at 1 µg/ml

Lysates/proteins at 10 µl per lane.


Observed band size : 48 kDa (why is the actual band size different from the predicted?)
Additional bands at : 65 kDa. We are unsure as to the identity of these extra bands.

MEL-1 and MEL-2 Human Embryonic Stem Cell Lysates (ab27198 and ab27196), IOUD2 Oct4-expressing Mouse Embryonic Stem Cell Lysate (ab272020) and F9 Mouse Embryonic Carcinoma Cell Lysate (ab27193) all contain Oct4 protein. As expected, a band corresponding to Oct4 was detected in all four of the lysates by Westen Blot using anti-Oct4 antibody ab19857. Each of the Oct4 bands was specifically blocked using the immunising peptide of ab19857. The origin of the additional 65 kDa band detected in Lane 3 is unknown and may be a non-specific band that is recognised, in addition to Oct4, by ab19857 in mouse embryonic stem cells.

 

Product image  Immunocytochemistry/ Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)

Immunocytochemistry/ Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)

ICC/IF image of ab19857 stained mouse embryonic stem cells. The cells were 4% formalin fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab19857, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).

 

Product image  Immunocytochemistry/ Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)

Immunocytochemistry/ Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)

ICC/IF image of ab19857 stained Mouse Embryonic Stem cells. The cells were PFA fixed (4% PFA, 20 min) and incubated with the antibody (ab19857, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue).

The large nuclei of the Feeder cells can be seen in the image; ab19857 does not localise to these nuclei. ab19857 can be seen localising to the much smaller nuclei of the Mouse Embryonic Stem cells.

 

Product image  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)

Mouse E6 - E6.5 embryos were sectioned and then fixed in paraformaldehyde. Antigen retrieval was performed using citric acid and the sections permeablized and blocked for 1 hour in serum. Embryos were stained with ab19857 (1/2500) in 10% serum, 1% ovalbumin in PBSTween for 14 hours at 4C. They were then washed and stained with a goat anti-rabbit biotin conjugated antibody (1/2000) and developed using ABC and then DAB for color. Staining is within the epiblast as expected.

See Abreview

 

Product image  Western blot

Western blot

All lanes : Oct4 antibody - ChIP Grade (ab19857) at 1 µg/ml

Lane 1 : Human embryonic stem cell lysate
Lane 2 : Mouse embryonic stem cell lysate
Lane 3 : Mouse embryonic germ cell lysate
Lane 4 : Human embryonic stem cell lysate with Oct4 peptide (ab20650) at 1 µg/ml
Lane 5 : Mouse embryonic stem cell lysate with Oct4 peptide (ab20650) at 1 µg/ml
Lane 6 : Mouse embryonic germ cell lysate with Oct4 peptide (ab20650) at 1 µg/ml

Lysates/proteins at 20 µg per lane.


Predicted band size : 39 kDa
Observed band size : 43 kDa (why is the actual band size different from the predicted?)
Additional bands at : 55 kDa (possible glycosylated form,cross reactivity).

Anti-Oct4 antiibody ab19857 only detected a band corresponding to the expected size of Oct4 in human ES cell lysate. In mouse ES and EG cell lysates, ab19857 detected a band of approximately 55 kDa in addition to the expected 39 kDa Oct4 band.

 

References for Oct4 antibody - ChIP Grade (ab19857)

Specific publication references   This product has been used in: (two most recent references)

Kim HM et al. Obox4 regulates the expression of histone family genes and promotes differentiation of mouse embryonic stem cells. FEBS Lett 584:605-11 (2010). WB; Mouse. PubMed: 20004198

Prasad SM et al. Continuous hypoxic culturing maintains activation of Notch and allows long-term propagation of human embryonic stem cells without spontaneous differentiation. Cell Prolif 42:63-74 (2009). ICC/IF; Human. PubMed: 19143764


See all 20 publication references for this product.


If you publish research using ab19857 please let us know so that we can cite the reference on this datasheet.

 

Search PubMed (MEDLINE) for references to Oct4

 

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Oct4 antibody - ChIP Grade - related products:

Products for ChIP

ChIP Related Products

Products for Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Slides

Products for Western blot

Loading controls

Lysates

 

Immunizing Peptide (Blocking) for Oct4 antibody - ChIP Grade

ab20650

  

Oct4 peptide

 

Immunizing Peptide for Oct4 antibody - ChIP Grade

 

Compatible Secondaries for Oct4 antibody - ChIP Grade

ab6564

  

Cy5 ® antibody

ab6717

  

FITC antibody

ab6720

  

Biotin antibody

ab6721

  

HRP antibody

ab6722

  

Alkaline Phosphatase antibody

ab6793

  

Texas Red ® antibody

ab7007

  

Phycoerythrin antibody

ab7019

  

Agarose antibody

ab27234

  

10nm Gold antibody

ab27235

  

5nm Gold antibody

ab27236

  

15nm Gold antibody

ab27237

  

20nm Gold antibody

ab27238

  

30nm Gold antibody

ab27239

  

10nm Gold antibody

ab27240

  

5nm Gold antibody

ab31193

  

Duck polyclonal to Rabbit IgG

ab60314

  

Chromeo™ 488 antibody

ab60315

  

Chromeo™ 494 antibody

ab60317

  

Chromeo™ 546 antibody

ab60319

  

Chromeo™ 642 antibody

 

Other Oct4 Antibodies

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