POU Class 5 Homeobox 1 (POU5F1) (AA 252-372) antibody

Details for Product No. ABIN968417
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Synonyms OCT3, OCT4, OTF-3, OTF3, OTF4, Oct-3, Oct-4, NF-A3, Oct-3/4, Oct3, Oct3/4, Oct4, Otf-3, Otf-4, Otf3, Otf3-rs7, Otf3g, Otf4, POU5F1, POU5F1L
AA 252-372
(17), (16), (11), (7), (4), (4), (4), (3), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Mouse (Murine)
(203), (87), (46), (33), (30), (28), (10), (5), (3), (3), (2), (2), (2), (1), (1)
(142), (65), (12), (10)
Clonality (Clone)
Monoclonal ()
(4), (4), (4), (4), (4), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1)
Western Blotting (WB), BioImaging (BI), Immunofluorescence (IF)
(165), (76), (47), (32), (31), (26), (20), (20), (5), (5), (4), (3), (3), (1), (1)
Pubmed 6 references available
Catalog no. ABIN968417
Quantity 50 µg
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Immunogen Mouse Oct3 Recombinant Protein
Clone DG10
Isotype IgG1
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 Purified from tissue culture supernatant or ascites by affinity chromatography.
Purity Purified
Alternative Name Oct-3
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
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.

Related Products: ABIN968631, ABIN967389

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
Preservative Sodium azide
Storage -20 °C
Supplier Images
anti-POU Class 5 Homeobox 1 (POU5F1) (AA 252-372) antibody Western Blot analysis of Oct3/4 in human and mouse ES cell lines. Lysates from H9 human ES cells (WiCell, Madison, WI, left blot) and ES-E14TG2a mouse ES cells (ATCC CRL-1821, right blot) and were probed with Purified Mouse anti-Oct3/4 monoclonal antibody at titrations of 2.0 (lanes 1), 1.0 (lanes 2), and 0.5 µg/ml (lanes 3). Oct3/4 is identified as a band of 46 kDa in the human and mouse ES cells.
The H9 cells were cultured on a mitomycin C-treated mouse embryonic fibroblast feeder layer [MEF (CF-1), ATCC SCRC-1040] that maintains the undifferentiated state of the ES cells. The lysate was made from a mixture of the 2 cell types, the majority of which were H9 cells.
anti-POU Class 5 Homeobox 1 (POU5F1) (AA 252-372) antibody (2) ES-E14TG2a mouse ES cells (ATCC CRL-1821)
anti-POU Class 5 Homeobox 1 (POU5F1) (AA 252-372) antibody (3) Bottom Row: Immunofluorescent staining of human and mouse ES cell lines. The H9 cell line (First Panel) and ES-E14TG2a cells (Second Panel) were cultured, fixed, permeabilized, and stained with Purified Oct3/4 monoclonal antibody (pseudo-colored green) according to the Recommended Assay Procedure. The second-step reagent was Alexa Fluor® 555 goat anti-mouse Ig (Invitrogen) and counter-staining was with Hoechst 33342 (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer using a 10X objective (H9) or 20X objective (E14) and merged using BD Attovision™ software. Permeabilization using BD Perm/Wash(TM) was used with this antibody, Triton™ X-100 or cold methanol will also work.
Product cited in: Okamoto, Okazawa, Okuda et al.: "A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells." in: Cell, Vol. 60, Issue 3, pp. 461-72, 1990 (PubMed).

Yuan, Corbi, Basilico et al.: "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 et al.: "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).

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).

Nishimoto, Fukushima, Okuda et al.: "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).

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

Hosts (142), (65), (12), (10)
Reactivities (203), (87), (46), (33), (30), (28), (10), (5), (3), (3), (2), (2), (2), (1), (1)
Applications (165), (76), (47), (32), (31), (26), (20), (20), (5), (5), (4), (3), (3), (1), (1)
Conjugates (4), (4), (4), (4), (4), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1)
Epitopes (17), (16), (11), (7), (4), (4), (4), (3), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
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