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    oct4-antibody-ab18976.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families Developmental Families OCT
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Anti-Oct4 antibody (ab18976)

  • Datasheet
  • SDS
Reviews (16)Q&A (10)References (180)

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Western blot - Anti-Oct4 antibody (ab18976)

    Key features and details

    • Rabbit polyclonal to Oct4
    • Suitable for: WB
    • Reacts with: Rat
    • Isotype: IgG

    Get better batch-to-batch reproducibility with a recombinant antibody

    Product image
    Anti-Oct4 antibody [EPR17980] (ab200834)
    • Research with confidence – consistent and reproducible results with every batch
    • Long-term and scalable supply – powered by recombinant technology for fast production
    • Success from the first experiment – confirmed specificity through extensive validation
    • Ethical standards compliant – production is animal-free

    Overview

    • Product name

      Anti-Oct4 antibody
      See all Oct4 primary antibodies
    • Description

      Rabbit polyclonal to Oct4
    • Host species

      Rabbit
    • Specificity

      The antibody reacts with 45 kDa Oct4 (also designated Oct3A) and in a less extent 33 kDa Oct3B from human, mouse, and rat samples.
    • Tested applications

      Suitable for: WBmore details
    • Species reactivity

      Reacts with: Rat
      Predicted to work with: Chimpanzee
    • Immunogen

      Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

    • General notes

      This product is manufactured by BioVision, an Abcam company and was previously called 3576 Oct-4 Antibody. 3576-100 is the same size as the 100 µg size of ab18976.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid
    • Storage instructions

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

      Preservative: 0.01% Thimerosal (merthiolate)
      Constituents: 69% PBS, 30% Glycerol (glycerin, glycerine), 0.5% BSA
    • Concentration information loading...
    • Purity

      Protein A purified
    • Purification notes

      Affinity Purified
    • Clonality

      Polyclonal
    • Isotype

      IgG
    • Research areas

      • Epigenetics and Nuclear Signaling
      • Transcription
      • Domain Families
      • Developmental Families
      • OCT
      • Stem Cells
      • Embryonic Stem Cells
      • Intracellular
      • Stem Cells
      • Germline Stem Cells
      • Embryonic Germ Cells
      • Developmental Biology
      • Reproduction
      • Germ cell markers
      • Developmental Biology
      • Embryogenesis
      • Embryonic stem cells
      • Surface molecules

    Associated products

    • Compatible Secondaries

      • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
      • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Isotype control

      • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
    • Recombinant Protein

      • Recombinant human Oct4 protein (ab169842)

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab18976 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    WB (5)
    Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 45 kDa.
    Notes
    WB
    Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 45 kDa.

    Target

    • Function

      Transcription factor that binds to the octamer motif (5'-ATTTGCAT-3'). Forms a trimeric complex with SOX2 on DNA and controls the expression of a number of genes involved in embryonic development such as YES1, FGF4, UTF1 and ZFP206. Critical for early embryogenesis and for embryonic stem cell pluripotency.
    • Tissue specificity

      Expressed in developing brain. Highest levels found in specific cell layers of the cortex, the olfactory bulb, the hippocampus and the cerebellum. Low levels of expression in adult tissues.
    • Sequence similarities

      Belongs to the POU transcription factor family. Class-5 subfamily.
      Contains 1 homeobox DNA-binding domain.
      Contains 1 POU-specific domain.
    • Developmental stage

      Highly expressed in undifferentiated embryonic stem cells and expression decreases gradually after embryoid body (EB) formation.
    • Domain

      The POU-specific domain mediates interaction with PKM2.
    • Post-translational
      modifications

      Sumoylation enhances the protein stability, DNA binding and transactivation activity. Sumoylation is required for enhanced YES1 expression.
      Ubiquitinated; undergoes 'Lys-63'-linked polyubiquitination by WWP2 leading to proteasomal degradation.
    • Cellular localization

      Nucleus. Expressed in a diffuse and slightly punctuate pattern.
    • Target information above from: UniProt accession Q01860 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 744263 Chimpanzee
      • Entrez Gene: 294562 Rat
      • SwissProt: Q7YR49 Chimpanzee
      • Alternative names

        • Octamer binding transcription factor 4 antibody
        • MGC22487 antibody
        • Oct 3 antibody
        • Oct 4 antibody
        • Oct-3 antibody
        • Oct-4 antibody
        • OCT3 antibody
        • Oct4 antibody
        • Octamer binding protein 3 antibody
        • Octamer binding protein 4 antibody
        • Octamer binding transcription factor 3 antibody
        • Octamer-binding protein 3 antibody
        • Octamer-binding protein 4 antibody
        • Octamer-binding transcription factor 3 antibody
        • OTF 3 antibody
        • OTF 4 antibody
        • OTF-3 antibody
        • OTF3 antibody
        • OTF4 antibody
        • PO5F1_HUMAN antibody
        • POU class 5 homeobox 1 antibody
        • POU domain class 5 transcription factor 1 antibody
        • POU domain transcription factor OCT4 antibody
        • POU domain, class 5, transcription factor 1 antibody
        • POU-type homeodomain-containing DNA-binding protein antibody
        • POU5F1 antibody
        see all

      Images

      • Western blot - Anti-Oct4 antibody (ab18976)
        Western blot - Anti-Oct4 antibody (ab18976)
        Anti-Oct4 antibody (ab18976) at 0.5 µg/ml + Lysates prepared from rat kidney tissue.

        Predicted band size: 45 kDa



        The antibody reacts with 45 kDa Oct4 (also designated Oct-3A) and in a less extent 33 kDa Oct-3B from human, mouse, and rat samples.

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (180)

      Publishing research using ab18976? Please let us know so that we can cite the reference in this datasheet.

      ab18976 has been referenced in 180 publications.

      • Huang Y  et al. The role of Chito-oligosaccharide in regulating ovarian germ stem cells function and restoring ovarian function in chemotherapy mice. Reprod Biol Endocrinol 19:14 (2021). PubMed: 33494759
      • Gong Y  et al. Transplantation of human induced pluripotent stem cell-derived neural crest cells for corneal endothelial regeneration. Stem Cell Res Ther 12:214 (2021). PubMed: 33781330
      • Yoon C  et al. PI3K/Akt pathway and Nanog maintain cancer stem cells in sarcomas. Oncogenesis 10:12 (2021). PubMed: 33468992
      • Loehr AR  et al. Targeting Cancer Stem Cells with Differentiation Agents as an Alternative to Genotoxic Chemotherapy for the Treatment of Malignant Testicular Germ Cell Tumors. Cancers (Basel) 13:N/A (2021). PubMed: 33922599
      • Silva DRS  et al. Characterization of rat liver bud-derived cells. Tissue Cell 71:101510 (2021). PubMed: 33721789
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-10 of 26 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Rat Cell (H9c2 rat cardiomyocytes)
      Permeabilization
      Yes - 0.01% TritonX-100
      Specification
      H9c2 rat cardiomyocytes
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
      Fixative
      Paraformaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Mar 18 2020

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Mouse Cell (Cardiomyocytes)
      Permeabilization
      Yes - 0.01% TritonX-100
      Specification
      Cardiomyocytes
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
      Fixative
      Paraformaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Mar 09 2020

      Western blot abreview for Anti-Oct4 antibody

      Average
      Abreviews
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (C42B Prostate cancer cell line)
      Gel Running Conditions
      Reduced Denaturing
      Loading amount
      20 µg
      Specification
      C42B Prostate cancer cell line
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      DR. Dimitra Kalamida

      Verified customer

      Submitted Dec 25 2019

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Pig Cell (Swine adipose-derived mesenchymal stem cells induc)
      Permeabilization
      Yes - 0.1 % Triton X-100
      Specification
      Swine adipose-derived mesenchymal stem cells induc
      Blocking step
      BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 23°C
      Fixative
      Formaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Dominic Schomberg

      Verified customer

      Submitted Apr 24 2019

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (PC3 Prostate cancer cells)
      Permeabilization
      Yes - 0.1% Triton X-100
      Specification
      PC3 Prostate cancer cells
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
      Fixative
      Paraformaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      DR. Dimitra Kalamida

      Verified customer

      Submitted Mar 21 2019

      Immunocytochemistry abreview for Anti-Oct4 antibody

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry
      Sample
      Mouse Cultured Cells (Neural stem cells)
      Permeabilization
      No
      Specification
      Neural stem cells
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
      Fixative
      Paraformaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Kyungjoo Seong

      Verified customer

      Submitted Oct 31 2016

      Flow Cytometry abreview for Anti-Oct4 antibody

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Flow Cytometry
      Sample
      Human Cell (H9 hESCs)
      Permeabilization
      Yes - 0.25% Triton X-100 in DPBS
      Gating Strategy
      Fibroblasts (white)
      Specification
      H9 hESCs
      Preparation
      Cell harvesting/tissue preparation method: Accutase
      Sample buffer: PBS
      Fixation
      Paraformaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Sep 24 2015

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Oct4 antibody

      Inconclusive
      Abreviews
      Abreviews
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Dog Tissue sections (Fetal Brain)
      Antigen retrieval step
      Other
      Permeabilization
      No
      Specification
      Fetal Brain
      Blocking step
      Dako protein block as blocking agent for 10 minute(s) · Concentration: .25% · Temperature: 20°C
      Fixative
      Formaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      MS. Erica Sloma

      Verified customer

      Submitted Oct 09 2014

      Question

      Dear Tech Support Team,



      Please read below the details of customer's complaint:



      Antibody code: ab18976

      General Information:

      Antibody storage conditions (temperature/reconstitution etc) (-20).

      Description of the problem (high background, low signal, non-specific staining etc.) no staining in ImmunoFluorescence staning.

      Sample (Species/Tissue/Cell Type/Cell Line etc.) Human & mouse cells & tissue

      Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.) Cells: methanol, Human tissue: formaldehyde, Mouse tissue: bouin.

      Antigen retrieval (Enzymatic method, Heat mediated technique etc.) Urea

      Permeabilization step

      Blocking conditions (Buffer/time period, Blocking agent etc.) Fetal Calf Serum in PBSX1 for 40mins

      Primary Antibody (Diluent/Dilution/Incubation time, Wash step)1:100 and 1:200 in blocking buffer for over night at 4C and washes with PBSX1 for 3 times, 5mins each.

      Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Fluorescent Anti Rabbit-1:1200 in blocking buffer for 40mins at room tempreture and washes in PBSX1 for 3 times, 10mins each.

      Detection method fluorescent microscope

      Positive and negative controls used (please specify) negative control: tissue/cells incubated with the secondary antibody without the first one.

      Positive control: tissues/cells stained with another antibody that works excellent (Sall-4 of abcam: ab

      Optimization attempts (problem solving):

      How many times have you tried the IHC? Our lab: since many years. Me: 2 years

      Have you run a "No Primary" control? Yes

      Do you obtain the same results every time? Yes

      What steps have you altered? All the steps are the same.



      The attached images:

      http://mail.zotal.co.il/exchange/Alla/Inbox/%D7%A6%D7%91%D7%99%D7%A2%D7%AA%20OCT%20%D7%91%D7%A8%D7%A7%D7%9E%D7%AA%20HUMAN.EML/1_multipart_xF8FF_3_Merged-30%2B31.jpg/C58EA28C-18C0-4a97-9AF2-036E93DDAFB3/Merged-30%2B31.jpg?attach=1 and http://mail.zotal.co.il/exchange/Alla/Inbox/%D7%A6%D7%91%D7%99%D7%A2%D7%AA%20OCT%20%D7%91%D7%A8%D7%A7%D7%9E%D7%AA%20HUMAN.EML/1_multipart_xF8FF_2_Merged-27%2B28.jpg/C58EA28C-18C0-4a97-9AF2-036E93DDAFB3/Merged-27%2B28.jpg?attach=1 - human sections, same dilutions, looks like the negative control



      All other attached images - mouse sections, 1/200 and 1/100, looks like the negative control



      Please advise.



      Thanks in advance.

      Have a nice week,

      Read More

      Abcam community

      Verified customer

      Asked on Nov 19 2012

      Answer

      Thank you and your customer for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody. The details your customer has kindly provided will enable us to investigate this case for your customer and this is also helpful in our records for monitoring of quality.
      Although the other antibody has worked well using this procedure, individual antibodies will often require optimization especially if the target is different. I would like to offer some suggestions to help optimise the results from ab18976.
      1.) I can recommend to add a permeabilization step to the protocol. Oct4 is in the nucleus. The following detergents can be used: Triton or NP-40 Use 0.1 to 0.2% in PBS for 10 min only. These will partially dissolve the nuclear membrane and are therefore very suitable for nuclear antigen staining.
      2.) Oct4 is expressed in developing brain. Highest levels are found in specific cell layers of the cortex, the olfactory bulb, the hippocampus and the cerebellum. Highly expressed in undifferentiated embryonic stem cells and expression decreases gradually after embryoid body (EB) formation. Since there is only a low level of expression in adult tissues, I suggest to run a positive control.
      I would also appreciate if your customer can confirm some details of the procedure.
      1.) What tissues and cells were used?
      2.) What does it urea mean as antigen retrieval step?
      3.) What cells are shown in the images?
      We are happy to offer this technical support. In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.
      I hope this information is helpful, thank you and your customer for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

      Read More

      Abcam Scientific Support

      Answered on Nov 19 2012

      Question

      Dear Sir / Madam,
      One of the biggest customer in stem cell lab would like to have an
      antibody that is specific against Oct 4, either variant A or variant B
      to distinguish between them. In literature, oct 4 antibodies are not
      able to bind to specific variants.
      What do you recommend?
      Many Thanks and Best Regards,

      Read More

      Abcam community

      Verified customer

      Asked on Jul 13 2012

      Answer

      Thank you for your enquiry and your interest.

      I can confirm that two isoforms of Oct 4 have been identified, and the exact amino acid sequences (aa) can be found at this website: http://www.uniprot.org/uniprot/Q01860

      - Isoform A: Q01860-1 (1-360 aa)

      - Isoform B: Q01860-2 (1-265 aa)

      ab109183 - this antibody was raised against a peptide corresponding to the region between 265-360 aa, soit is very likely to recognize Isoform A.

      ab18976 - the immunogen selected was around region 1-14 aa, thus it would recognize potentially both Isofm A and B.

      If you need any further assistance in the future, please do not hesitate to contact me.

      Read More

      Abcam Scientific Support

      Answered on Jul 13 2012

      1-10 of 26 Abreviews or Q&A

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