OCT4 antibody (AA 252-372)

Catalog No. ABIN968417

Product Details anti-OCT4 Antibody

Target: OCT4 (POU5F1) (POU Class 5 Homeobox 1 (POU5F1))

Binding Specificity: AA 252-372

Reactivity: Human, Mouse

Host: Mouse

Clonality: Monoclonal

Conjugate: This OCT4 antibody is un-conjugated

Application: Western Blotting (WB), Immunofluorescence (IF), BioImaging (BI)

Cross-Reactivity: Human

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Please refer to us for technical protocols.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Mouse Oct3 aa. 252-372 Recombinant Protein

Clone: 40-Oct

Isotype: IgG1 kappa

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at an appropriate cell density in an 96-well Imaging Plate , and culture overnight to 48 hours.
2. Remove the culture medium from the wells, wash the wells twice with 100 µl of 1x PBS, and fix the cells by adding 100 µl of fresh 3.7% Formaldehyde in PBS or fixation buffer to each well and incubating for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and wash the wells twice with 100 µl of 1x PBS.
4. Permeabilize the cells by adding 100 µl of 1× Perm/Wash™ buffer to each well and incubating for 30 minutes at RT.
5. Remove the permeabilizer, and wash the wells twice with 100 µl of 1x PBS.
6. Dilute the antibody in Perm/Wash™ buffer, and stain the cells by adding 50 µl of the diluted antibody to each well and incubating for 1 hour at RT.
7. Remove the diluted antibody, and wash the wells three times with 100 µl of 1x PBS.
8. Remove the PBS, dilute the second-step reagent in Perm/Wash™ buffer, and stain the cells by adding 50 µl of the diluted second-step reagent to each well and incubating for 1 hour at RT.
9. Remove the diluted second-step reagent, and wash the wells twice with 100 µl of 1x PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 100 µl of a 2 µg/ml solution of Hoechst 33342 in 1x PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN968631, ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20° C.

References for anti-OCT4 antibody (ABIN968417)

Pan, Thomson: "Nanog and transcriptional networks in embryonic stem cell pluripotency." in: Cell research, Vol. 17, Issue 1, pp. 42-9, (2007) (PubMed).

Nishimoto, Fukushima, Okuda, Muramatsu: "The gene for the embryonic stem cell coactivator UTF1 carries a regulatory element which selectively interacts with a complex composed of Oct-3/4 and Sox-2." in: Molecular and cellular biology, Vol. 19, Issue 8, pp. 5453-65, (1999) (PubMed).

Vigano, Staudt: "Transcriptional activation by Oct-3: evidence for a specific role of the POU-specific domain in mediating functional interaction with Oct-1." in: Nucleic acids research, Vol. 24, Issue 11, pp. 2112-8, (1996) (PubMed).

Yuan, Corbi, Basilico, Dailey: "Developmental-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3." in: Genes & development, Vol. 9, Issue 21, pp. 2635-45, (1995) (PubMed).

Rosfjord, Scholtz, Lewis, Rizzino: "Phosphorylation and DNA binding of the octamer binding transcription factor Oct-3." in: Biochemical and biophysical research communications, Vol. 212, Issue 3, pp. 847-53, (1995) (PubMed).

Okamoto, Okazawa, Okuda, Sakai, Muramatsu, Hamada: "A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells." in: Cell, Vol. 60, Issue 3, pp. 461-72, (1990) (PubMed).

Target Details for OCT4

Target: OCT4 (POU5F1) (POU Class 5 Homeobox 1 (POU5F1))

Alternative Name: Oct-3 (POU5F1 Products)

Synonyms: OCT3 antibody, OCT4 antibody, OTF-3 antibody, OTF3 antibody, OTF4 antibody, Oct-3 antibody, Oct-4 antibody, POU5F1 antibody, gp9 antibody, gp-9 antibody, oct4 antibody, pou2 antibody, POU-2 antibody, cb197 antibody, spg/pou2 antibody, chunp6868 antibody, wu:fd18d06 antibody, NF-A3 antibody, Oct-3/4 antibody, Oct3 antibody, Oct3/4 antibody, Oct4 antibody, Otf-3 antibody, Otf-4 antibody, Otf3 antibody, Otf3-rs7 antibody, Otf3g antibody, Otf4 antibody, POU class 5 homeobox 1 antibody, POU domain, class 5, transcription factor 3 antibody, POU domain, class 5, transcription factor 1 antibody, POU5F1 antibody, pou5f3 antibody, Pou5f1 antibody

Background: Development of a multicellular organism from a single fertilized cell is regulated by the coordinated activity of DNA transcription factors. Oct3/4, a member of the POU family of transcription factors, functions in pluripotent cells of early embryonic stem cell (ES) lines and embryonal carcinomas (EC). Other members of the POU family include Oct1, Oct2, Pit-1, and unc-86. The POU domain, a 150-amino acid region that determines binding specificity, is conserved among these proteins and consists of 3 subdomains: POU-specific A and B subdomains and a homeobox-like subdomain. Oct3/4 is expressed in undifferentiated cells, but is lost as cells are induced to differentiate. Oct3/4 is not expressed in adult tissues. The interaction of Oct3/4 with SOX2, another embryonic transcription factor, produces an active complex that regulates expression of genes such as Nanog, UTF1, and FGF4. Although Oct3/4 is specifically phosphorylated on serine residues, this modification is not required for DNA binding, but may affect its transactivation potential. Thus, Oct3/4 is a transcription factor that plays an important role in determining early steps of embryogenesis and differentiation.
Synonyms: Oct3, OTF3, Oct4, OTF4, POU5F1

Molecular Weight: 46 kDa

Pathways: Stem Cell Maintenance