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    oct4-antibody-ab19857.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families Developmental Families OCT
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Anti-Oct4 antibody (ab19857)

  • Datasheet
  • SDS
Reviews (44)Q&A (32)References (459)

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Immunocytochemistry - Anti-Oct4 antibody (ab19857)
  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (ab19857)
  • Western blot - Anti-Oct4 antibody (ab19857)
  • Immunoprecipitation - Anti-Oct4 antibody (ab19857)
  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (ab19857)
  • Western blot - Anti-Oct4 antibody (ab19857)
  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (ab19857)

Key features and details

  • Rabbit polyclonal to Oct4
  • Suitable for: IP, ICC/IF, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

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

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

    Previous batches of ab19857 have worked well in IHC, but recent batches of this antibody are showing non-specific staining on day 6 mouse testis in our lab (as of Nov 2016).
  • Tested applications

    Suitable for: IP, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Sheep, Rhesus monkey
  • Immunogen

    Synthetic peptide corresponding to Human Oct4 aa 300 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab20650)

  • General notes

    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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: 99% PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help
  • 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

  • Assay kits

    • Oct4 Transcription Factor Assay Kit (Colorimetric) (ab207224)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Immunizing Peptide (Blocking)

    • Human Oct4 peptide (ab20650)
  • Isotype control

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

    • Recombinant human Oct4 protein (ab169842)
  • Related Buffer

    • trans-Retinoic acid, Stem cell differentiator (ab120728)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab19857 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
IP
Use at an assay dependent concentration.
ICC/IF (11)
Use a concentration of 1 - 5 µg/ml.
WB (10)
Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 39 kDa).
Notes
IP
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 1 - 5 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 39 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: 5460 Human
    • Entrez Gene: 18999 Mouse
    • Entrez Gene: 294562 Rat
    • Entrez Gene: 714760 Rhesus monkey
    • Omim: 164177 Human
    • SwissProt: Q01860 Human
    • SwissProt: P20263 Mouse
    • SwissProt: Q5TM49 Rhesus monkey
    • Unigene: 249184 Human
    • Unigene: 632482 Human
    • Unigene: 646545 Human
    • Unigene: 17031 Mouse
    see all
  • 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

  • Immunocytochemistry - Anti-Oct4 antibody (ab19857)
    Immunocytochemistry - Anti-Oct4 antibody (ab19857)

    ab19857 staining Oct4 in mES cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab19857 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Also suitable in cells fixed with 4% paraformaldehyde (10 min).

    Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (ab19857)
    Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (ab19857)

    ab19857 staining Oct4 in F9 cells treated with trans-retinoic acid (ab120728) The cells were incubated at 37°C for 2 days in media containing different concentrations of ab120728 (trans-retinoic acid) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab19857 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1:250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • Western blot - Anti-Oct4 antibody (ab19857)
    Western blot - Anti-Oct4 antibody (ab19857)
    All lanes : Anti-Oct4 antibody (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 Human Oct4 peptide (ab20650) at 1 µg/ml
    Lane 6 : MEL-2 (Human embryonic stem cell, female cell line) Whole Cell Lysate (ab27196) with Human 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 Human Oct4 peptide (ab20650) at 1 µg/ml
    Lane 8 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate (ab27193) with Human Oct4 peptide (ab20650) at 1 µg/ml

    Lysates/proteins at 10 µl per lane.

    Predicted band size: 39 kDa
    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 (ab27202) 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.

  • Immunoprecipitation - Anti-Oct4 antibody (ab19857)
    Immunoprecipitation - Anti-Oct4 antibody (ab19857)

    Oct4 was immunoprecipitated using 0.5mg E14Tg2a whole cell extract, 5µg of Rabbit polyclonal to Oct4 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, E14Tg2a whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab19857.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 48kDa: Oct4.

  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (ab19857)
    Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (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-iT™ 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.

  • Western blot - Anti-Oct4 antibody (ab19857)
    Western blot - Anti-Oct4 antibody (ab19857)
    All lanes : Anti-Oct4 antibody (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 Human Oct4 peptide (ab20650)
    Lane 5 : Mouse embryonic stem cell lysate with Human Oct4 peptide (ab20650)
    Lane 6 : Mouse embryonic germ cell lysate with Human Oct4 peptide (ab20650)

    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), 55 kDa (possible cross reactivity)



    Anti-Oct4 antibody 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (ab19857)
    Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody (ab19857)This image is courtesy of Ludovic Vallier, University of Cambridge

    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 Oct-4-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.

Protocols

  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (459)

Publishing research using ab19857? Please let us know so that we can cite the reference in this datasheet.

ab19857 has been referenced in 459 publications.

  • Hernández-Ainsa C  et al. Development and characterization of cell models harbouring mtDNA deletions for in vitro study of Pearson syndrome. Dis Model Mech 15:N/A (2022). PubMed: 35191981
  • Kim JW  et al. Prognostic implication of SOX2 expression in small intestinal adenocarcinoma. Virchows Arch 478:1049-1060 (2021). PubMed: 33103210
  • Ugurlu-Çimen D  et al. AF10 (MLLT10) prevents somatic cell reprogramming through regulation of DOT1L-mediated H3K79 methylation. Epigenetics Chromatin 14:32 (2021). PubMed: 34215314
  • Shan W  et al. Polycomb group protein Bmi1 is required for the neuronal differentiation of mouse induced pluripotent stem cells. Exp Ther Med 21:619 (2021). PubMed: 33936276
  • Nathansen J  et al. Oct4 confers stemness and radioresistance to head and neck squamous cell carcinoma by regulating the homologous recombination factors PSMC3IP and RAD54L. Oncogene 40:4214-4228 (2021). PubMed: 34079088
View all Publications for this product

Customer reviews and Q&As

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1-10 of 76 Abreviews or Q&A

Western blot abreview for Anti-Oct4 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (pancreas, lung)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
pancreas, lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°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.

Abcam user community

Verified customer

Submitted Aug 30 2021

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

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Ph:6.0
Permeabilization
No
Specification
Pancreas
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°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 Nov 01 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human iPSC colonies)
Permeabilization
Yes - 0.25% Triton X-100
Specification
human iPSC colonies
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°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.

Abcam user community

Verified customer

Submitted Jul 15 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (neural precursors)
Permeabilization
Yes - tritonX 0.05%
Specification
neural precursors
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°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 Oct 15 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (IPSC)
Permeabilization
Yes - 0.3% Triton X-100
Specification
IPSC
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°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.

Jacob Garza

Verified customer

Submitted Jul 27 2018

ChIP abreview for Anti-Oct4 antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (hESC)
Negative control
gene desert
Specification
hESC
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 8 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% PFA
Positive control
upstream and promoter
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.

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Verified customer

Submitted Aug 31 2017

IHC - Wholemount abreview for Anti-Oct4 antibody - ChIP Grade

Poor
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Tissue (mouse P6 testis)
Specification
mouse P6 testis
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.

Jin Yue Liao

Verified customer

Submitted Aug 18 2017

Mass Cytometry abreview for Anti-Oct4 antibody

Inconclusive
Abreviews
Abreviews
Application
Mass Cytometry
Sample
Mouse Cell (lung)
Specification
lung
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.

Irit Milman Krentsis

Verified customer

Submitted Feb 06 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (induced pluripotent cells)
Permeabilization
Yes - 0.5% TX100
Specification
induced pluripotent cells
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°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.

HERR DR. Vladimir Milenkovic

Verified customer

Submitted Nov 04 2016

Immunohistochemistry (Frozen sections) abreview for Anti-Oct4 antibody - ChIP Grade

Poor
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, postnatal day 6)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, postnatal day 6
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°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.

MR. Bryan Niedenberger

Verified customer

Submitted May 06 2016

1-10 of 76 Abreviews or Q&A

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