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    products/primary-antibodies/oct4-antibody-epr17980-ab200834.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families Developmental Families OCT
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RecombinantRabMAb

Recombinant Anti-Oct4 antibody [EPR17980] (ab200834)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (4) Submit a question References (11)

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Western blot - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Western blot - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Flow Cytometry (Intracellular) - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Immunoprecipitation - Anti-Oct4 antibody [EPR17980] (ab200834)
  • ChIP-sequencing - Anti-Oct4 antibody [EPR17980] (ab200834)
  • ChIC/CUT&RUN sequencing - Anti-Oct4 antibody [EPR17980] (ab200834)
  • Anti-Oct4 antibody [EPR17980] (ab200834)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17980] to Oct4
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seq, ChIP-sequencing
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free PE

You may also be interested in

Biochemical
Product image
AG 490, JAK2/3 inhibitor (ab120950)
Protein
Recombinant human Oct4 protein (ab169842)
Primary
Product image
Alexa Fluor® 488 Anti-Oct4 antibody [EPR17980] (ab208907)

View more associated products

Overview

  • Product name

    Anti-Oct4 antibody [EPR17980]
    See all Oct4 primary antibodies
  • Description

    Rabbit monoclonal [EPR17980] to Oct4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seq, ChIP-sequencingmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: NCCIT, Ntera-2/D1 and F9 whole cell lysates. IHC-P: Human dysgerminoma of ovary and spermatocytoma tissues. ICC/IF: NCCIT cells, F9 cells Flow Cyt (intra): F9 cells. IP: F9 whole cell lysate. ChIC/CUT&RUN-seq: NCCIT cells. ChIP-seq: NCCIT 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.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR17980
  • 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

  • Alternative Versions

    • Alexa Fluor® 488 Anti-Oct4 antibody [EPR17980] (ab208907)
    • Alexa Fluor® 647 Anti-Oct4 antibody [EPR17980] (ab209957)
    • PE Anti-Oct4 antibody [EPR17980] (ab213198)
    • Alexa Fluor® 594 Anti-Oct4 antibody [EPR17980] (ab217250)
    • Alexa Fluor® 555 Anti-Oct4 antibody [EPR17980] (ab217743)
    • Anti-Oct4 antibody [EPR17980] - BSA and Azide free (ab240358)
  • Assay kits

    • Oct4 Transcription Factor Assay Kit (Colorimetric) (ab207224)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human Oct4 protein (ab169842)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab200834 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
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 38 kDa).
IHC-P (1)
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF (1)
Use a concentration of 5 µg/ml.
IP
1/30.
Flow Cyt (Intra)
1/60.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ChIC/CUT&RUN-seq
Use at an assay dependent concentration.
ChIP-sequencing
Use at an assay dependent concentration.
Notes
WB
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 38 kDa).
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 5 µg/ml.
IP
1/30.
Flow Cyt (Intra)
1/60.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ChIC/CUT&RUN-seq
Use at an assay dependent concentration.
ChIP-sequencing
Use at an assay dependent concentration.

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
    • Omim: 164177 Human
    • SwissProt: Q01860 Human
    • SwissProt: P20263 Mouse
    • 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

  • Western blot - Anti-Oct4 antibody [EPR17980] (ab200834)
    Western blot - Anti-Oct4 antibody [EPR17980] (ab200834)
    All lanes : Anti-Oct4 antibody [EPR17980] (ab200834) at 1/10000 dilution

    Lane 1 : NCCIT (Human pluripotent embryonic carcinoma) whole cell lysate
    Lane 2 : NTera-2/D1 (Human malignant pluripotent embryonic carcinoma) whole cell lysate
    Lane 3 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
    Lane 4 : JAR (Human placenta choriocarcinoma cell line) whole cell lysate
    Lane 5 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 6 : Mouse testis 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: 38 kDa
    Observed band size: 45 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Low levels of expression in adult tissues.

    Blocking/Dilution Buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17980] (ab200834)
    Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17980] (ab200834)

    ab200834 staining Oct4 in F9 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 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 ab200834 at a 5μg/ml concentration and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1μg/ml concentration, followed by a further incubation at room temperature for 1h with an anti-rabbit AlexaFluor® 488 (ab150081) at 2 μg/ml (shown in green) and an anti-mouse AlexaFluor® 594 (ab150120) at 2 μg/ml (shown in pseudocolor red).  Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17980] (ab200834)
    Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17980] (ab200834)

    Immunocytochemistry/Immunofluorescence analysis of F9 (mouse embryonal carcinoma) labelling Oct4 with purified ab200834 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

  • Western blot - Anti-Oct4 antibody [EPR17980] (ab200834)
    Western blot - Anti-Oct4 antibody [EPR17980] (ab200834)
    All lanes : Anti-Oct4 antibody [EPR17980] (ab200834) at 1/1000 dilution

    Lane 1 : Human placenta lysate
    Lane 2 : Human fetal brain lysate
    Lane 3 : Human testis lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 38 kDa


    Exposure time: 3 minutes


    Low levels of expression in adult tissues

    Blocking/Dilution Buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)

    Immunohistochemical analysis of paraffin-embedded Human dysgerminoma of ovary tissue labeling Oct4 with ab200834 at 1/500 dilution, followed Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weakly cytoplasmic staining on Human dysgerminoma of ovary tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only 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 Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)

    Immunohistochemical analysis of paraffin-embedded Human spermatocytoma tissue labeling Oct4 with ab200834 at 1/500 dilution, followed Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weakly cytoplasmic staining on Human spermatocytoma tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only 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 Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)

    Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Oct4 with ab200834 at 1/500 dilution, followed Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Human cerebral cortex tissue is a negative control for Oct4. Counter stained with Hematoxylin.

    Secondary antibody only 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 Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry (Intracellular) - Anti-Oct4 antibody [EPR17980] (ab200834)
    Flow Cytometry (Intracellular) - Anti-Oct4 antibody [EPR17980] (ab200834)

    Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed F9 (Mouse embyro testicular cancer cell line) cells labeling Oct4 with ab200834 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730;black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody. 

     

     

  • Immunoprecipitation - Anti-Oct4 antibody [EPR17980] (ab200834)
    Immunoprecipitation - Anti-Oct4 antibody [EPR17980] (ab200834)

    Oct4 was immunoprecipitated from 1mg of F9 (Mouse embyro testicular cancer cell line) whole cell lysate with ab200834 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab200834 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: F9 whole cell lysate 10 µg (Input). Lane 2: ab200834 IP in F9 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200834 in F9 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds

  • ChIP-sequencing - Anti-Oct4 antibody [EPR17980] (ab200834)
    ChIP-sequencing - Anti-Oct4 antibody [EPR17980] (ab200834)

    Chromatin was prepared from NCCIT (Human pluripotent embryonic carcinoma cell line) cells. ChIP was performed with 10^7 NCCIT cells and 8 µg of ab200834 [EPR17980]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
    Additional screenshots of mapped reads can be downloaded here.

  • ChIC/CUT&RUN sequencing - Anti-Oct4 antibody [EPR17980] (ab200834)
    ChIC/CUT&RUN sequencing - Anti-Oct4 antibody [EPR17980] (ab200834)

    CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373, 2.5 x 105 NCCIT (Human pluripotent embryonic carcinoma cell line) cells and 5 µg of ab200834 [EPR17980]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads.

    Additional screenshots of mapped reads can be downloaded here.

  • Anti-Oct4 antibody [EPR17980] (ab200834)
    Anti-Oct4 antibody [EPR17980] (ab200834)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (11)

Publishing research using ab200834? Please let us know so that we can cite the reference in this datasheet.

ab200834 has been referenced in 11 publications.

  • Yang X  et al. Downregulation of hsa_circ_0026123 suppresses ovarian cancer cell metastasis and proliferation through the miR-124-3p/EZH2 signaling pathway. Int J Mol Med 47:668-676 (2021). PubMed: 33416105
  • Li S  et al. Schisandrin B inhibits epithelial-mesenchymal transition and stemness of large-cell lung cancer cells and tumorigenesis in xenografts via inhibiting the NF-?B and p38 MAPK signaling pathways. Oncol Rep 45:N/A (2021). PubMed: 33907830
  • Wu Y  et al. LncRNA WDFY3-AS2 promotes cisplatin resistance and the cancer stem cell in ovarian cancer by regulating hsa-miR-139-5p/SDC4 axis. Cancer Cell Int 21:284 (2021). PubMed: 34051810
  • Zhang W  et al. Roles of Melatonin in Goat Hair Follicle Stem Cell Proliferation and Pluripotency Through Regulating the Wnt Signaling Pathway. Front Cell Dev Biol 9:686805 (2021). PubMed: 34150780
  • Chen M  et al. Chemokine CCL20 promotes the paclitaxel resistance of CD44+CD117+ cells via the Notch1 signaling pathway in ovarian cancer. Mol Med Rep 24:N/A (2021). PubMed: 34278466
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-4 of 4 Abreviews or Q&A

IHC - Wholemount abreview for Anti-Oct4 antibody [EPR17980]

Excellent
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Tissue (120h Blimp1-mVenus Stella-eCFP gastruloid)
Specification
120h Blimp1-mVenus Stella-eCFP gastruloid
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. Chris Cooke

Verified customer

Submitted Mar 09 2023

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Oct4 antibody [EPR17980]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (E6.5 mouse embryo)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9 and Citric Acid Buffer pH 6
Permeabilization
Yes - PBT
Specification
E6.5 mouse embryo
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 0.5% · 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.

Abcam user community

Verified customer

Submitted Jun 29 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Oct4 antibody [EPR17980]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hiPSC derived embryoid bodies)
Permeabilization
No
Specification
hiPSC derived embryoid bodies
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · 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.

Abcam user community

Verified customer

Submitted Mar 27 2019

Immunohistochemistry (Frozen sections) abreview for Anti-Oct4 antibody [EPR17980]

Good
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 Mar 04 2016

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