NANOG Antibody (OASA08018)

Catalog#: OASA08018

• Size: 0.1mg

• Description of Target: RABBIT ANTI HUMAN NANOG
• Gene Symbol: NANOG
• Alias Symbols: NANOG
• Protein Accession#: NP_079141.2
• Swissprot Id: Q9H9S0
• Host: Rabbit
• Protein Size (# AA): 305
• Specificity: NANOG
• Application: ELISA, WB
• Immunogen: The immunogen for anti-NANOG antibody: recombinant human nanog
• Product Format: Purified IgG - lyophilised
• Isotype: Polyclonal IgG
• Predicted Homology Based on Immunogen Sequence: Human
• Applications Info: ELISA : This antibody may be used in an indirect ELISA or as a capture reagent in a sandwich ELISA with This product B as the detection antibody.
  Preservative Stabilisers: None present
  
  Antiserum Preparation: Antiserum to human nanog were raised by repeated immunisation of rabbits with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
  Approx Protein Conc: IgG concentration 1.0mg/ml after reconstitution
  
  Buffer Solutions: Phosphate buffered saline pH7.2
• Reconstitution and Storage: Reconstitution: Reconstitute with 0.1ml distilled water
  Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Aviva Systems Biology recommend that the vial is gently mixed after reconstitution. For l

Product Protocol: NANOG antibody used to evaluate characterization of an acyl-coenzyme a binding protein predominantly expressed in human primitive progenitor cells (OASA08018)

Specificity: NANOG
Product Page: NANOG antibody (OASA08018)
Antibody Pair Available: OASA08018 Capture OASA08019 Detection
Experiment Type: Immunohistochemistry

Protocol:
After institutional review board approval and informed consent were secured, human term placentas were obtained from healthy females after cesarean section at Alta Bates Medical Center in Berkeley, CA. Only freshly obtained placentas, drained substantially of all cord blood by a conventional collection process, were used. They were infused with an anticoagulant and vasodilator solution (30 U/ml heparin and 1 mg of papaverin hydrochloride) at room temperature. The tissue was then fixed in 2.4% paraformaldehyde for 24 h, washed with PBS, pH 7.4, embedded in paraffin, and sectioned. Before immunostaining, paraffin sections were deparaffinized in xylene (Sigma), rehydrated in alcohol, and washed in PBS. Antigen retrieval was performed with protease K (20 U/ml in 10 mM Tris-HCl pH 7.5, 1 mM EDTA buffer) for 3 min at room temperature. Sections were then incubated with a blocking solution containing 2% goat serum, 3% fetal calf serum, 0.1% Tween-20, and 3% BSA in 4× saline-sodium citrate for 60 min at 37°C. They were then incubated with the primary antibody at a 1:100 dilution overnight at 4°C. Sections were washed, incubated with the blocking solution for 20 min, and then incubated with a secondary antibody at a 1:500 dilution for 60 min at 37°C. This antibody was labeled with either FITC or Alexa Fluor 633. Finally, sections were washed and mounted on slides with Gold Antifade reagent (Molecular Probes, Invitrogen, Carlsbad, CA). The following antibodies were used: mouse anti-human CD34 (No. 555820; BD Pharmingen, San Jose, CA), mouse anti-human CD31 (No. BM4047; Acris Antibodies GmbH), rabbit anti-human CD133 (No. ab16518; Abcam, Cambridge, MA), rat anti-human SSEA-3 (No. ab16286; Abcam), rabbit anti-human Oct-4 (No. ab19857; Abcam), and rabbit anti-human Nanog (No. ab21603; Abcam). Isotype anti-mouse and anti-rabbit antibodies were from Zymed (South San Francisco, CA). Secondary anti-mouse and anti-rabbit FITC- or Alexa Fluor 633-labeled antibodies were from Molecular Probes, Invitrogen.

Summary:
Eric et al. established several lines of plastic-adherent cells from human placentas. These cells were negative for CD45, CD34, and CD38 and propagated up to 100 doublings in cell culture. They were highly positive for Oct-4, Nanog, and SSEA-3, classical markers of human embryonic stem cells.

References:
1: Soupene E, Serikov V, Kuypers FA. Characterization of an acyl-coenzyme A binding protein predominantly expressed in human primitive progenitor cells. J Lipid Res. 2008 May;49(5):1103-12. Epub 2008 Feb 11. PubMed PMID: 18268358; PubMed Central PMCID: PMC2311440.