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Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)
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Western blot
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Immunocytochemistry/ Immunofluorescence - Oct4 antibody - Embryonic Stem Cell Marker (ab19857)
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Product Name
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Oct4 antibody - ChIP Grade
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See all Oct4 antibodies (14)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Oct4 - ChIP Grade
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human Oct4.
(Peptide available as ab20650.)
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Reacts with
(species key)
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Hu, Ms
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Specificity
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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.
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Tested applications
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ChIP, Flow Cyt, ICC/IF, IHC-P, WB
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Abreviews
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Application notes
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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.
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Positive control
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Human ES cell lysate
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Cellular localization
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Nuclear
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Research areas
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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
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Relevance
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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.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.10 - 0.90 mg/ml
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Storage instructions
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Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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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! |
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See below for Oct4 antibody - ChIP Grade images, references, products related to ab19857 and other tools.
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Oct4 antibody - ChIP Grade images:
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 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
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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.
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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).
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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.
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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
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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.
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References for Oct4 antibody - ChIP Grade (ab19857)
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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.
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Search PubMed (MEDLINE) for references to Oct4
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