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Human DPPA4 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF3674

R&D Systems, part of Bio-Techne
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AF3674
AF3674-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

CyTOF-ready, Immunocytochemistry, Intracellular Staining by Flow Cytometry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

Product Specifications

Immunogen

E. coli-derived recombinant human DPPA4
Met1-Glu304
Accession # Q7L190

Specificity

Detects human DPPA4 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant mouse DPPA4 is observed and less than 1% cross-reactivity with recombinant human DPPA5 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human DPPA4 Antibody

Detection of DPPA4 antibody in NTera-2 Human Cell Line antibody by Flow Cytometry.

Detection of DPPA4 in NTera-2 Human Cell Line by Flow Cytometry.

NTera-2 human testicular embryonic carcinoma cell line was stained with Sheep Anti-Human DPPA4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3674, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL010). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
DPPA4 antibody in Human iPSK3 cells by Immunocytochemistry (ICC).

DPPA4 in Human iPSK3 cells.

DPPA4 was detected in immersion fixed human plasmid-derived induced pluripotent stem cells (iPSK3) using Sheep Anti-Human DPPA4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3674) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Detection of Mouse DPPA4 by Immunocytochemistry/Immunofluorescence

Detection of Mouse DPPA4 by Immunocytochemistry/Immunofluorescence

Identification of USP26 as a negative regulator of pluripotency. a Schematic of experimental strategy for screening essential Usps for the generation of iPSCs. Dox-inducible OSKM-transgenic MEFs were plated, transfected with 48 individual mouse Usp shRNA lentiviral vectors to knockdown Usps, and stained for AP+ iPSC colonies after 12 days of Dox treatment. b Quantification of AP+ colonies after 12 days of OSKM induction in MEFs transduced with shRNA, targeting members of the mouse Usp family, p < 0.001 compared to control shRNA. c Quantification of AP+ colonies after 12 days of OSKM induction in MEFs transduced with mouse Usp26 shRNA, Usp20 shRNA, or control shRNA lentivirus, *p < 0.05 compared to control shRNA, **p < 0.01 compared to control shRNA. d Quantification of AP+ colonies after 12 days of OSKM induction in MEFs transduced with pLtet-O (tetracycline-inducible) mouse Usp26, Usp20, or empty vector (EV) lentivirus, **p < 0.01 compared to EV. e Images of AP staining of iPSC colonies after 12 days’ OSKM induction in MEFs transduced with control shRNA, Usp26 shRNA, EV, or pLtet-O lentivirus overexpressing Usp26. f Bright field (BF) and immunofluorescence microscopic images of Oct4, Nanog, and Dppa4 in Usp26 knockdown iPSCs. iPSC colonies were fixed, blocked, and stained with specific antibodies, followed by goat anti-mouse antibody-conjugated Texas Red. Nuclei were stained with DAPI. Scale bar, 100 µM. The data are presented as means ± SD from three independent experiments. b–d Two-way ANOVA for multiple comparisons Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28839133), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human DPPA4 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human plasmid-derived induced pluripotent stem cells (iPSK3)

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: NTera-2 human testicular embryonic carcinoma cell line fixed with paraformaldehyde and permeabilized with saponin

Western Blot

0.1 µg/mL
Sample: Recombinant Human DPPA4

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: DPPA4

DPPA4 is a nuclear SAP-domain protein whose expression is restricted to pluripotent cells, undifferentiated embryonal carcinomas and germ cell tumors. It is considered a marker for embryonic stem cells. Human DPPA4 shares 51% amino acid sequence homology with mouse DPPA4.

Long Name

Developmental Pluripotency Associated 4

Alternate Names

developmental pluripotency associated 4, developmental pluripotency-associated protein 4, FLJ10713,2410091M23Rik

Entrez Gene IDs

55211 (Human); 73693 (Mouse)

Gene Symbol

DPPA4

UniProt

Additional DPPA4 Products

Product Documents for Human DPPA4 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human DPPA4 Antibody

For research use only

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