POU Class 5 Homeobox 1 (POU5F1) antibody

Antigen: POU Class 5 Homeobox 1 (POU5F1)

Synonyms: OCT3, OCT4, OTF3, OTF4, MGC22487, POU5F1, POU5F1L

Clonality: Polyclonal

Host: Rabbit

Reactivity: Human

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN349596

Additional Information

Alternative name: Oct-4

Gene ID: NP_976034

Swiss-Prot: Q01860

Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region of human Oct-4 protein.

Format: Liquid (sterile filtered)

Description: Oct-4, also known as POU domain transcription factor, Octomer- binding transcription factor 3 and Oct-3, is expressed in human and mouse totipotent embryonic stem and germ cells. It may function in preventing these cells from differentiating. Each type of established mammalian pluripotent stem cell line (embryonal carcinoma (EC) cells, embryonic stem (ES) cells and embryonic germ (EG) cells) expresses Oct-4, which disappears rapidly when the cells differentiate. Most studies on Oct-4 have been performed on murine embryos. In human oocytes of various maturity and normally developing preimplantation/cleavage embryos, a variable Oct-4 expression pattern has been revealed, concomitant with a pure cytoplasmic localization of the protein. This variability in expression fades away during compaction, indicating embryonic Oct-4 expression, the protein appears in the nucleus, implying biological activity. Functional Oct-4 has been found in human amniotic fluid. As Oct-4 transcripts have also been found in various differentiated cells, the presence of Oct-4 transcripts or proteins may not be sufficient for identifying undifferentiated cell lines in humans. It may also be important to examine the localization of Oct-4 proteins within a cell. Oct-4 has been shown to interact with Nanog and with proteins from multiple repression complexes, including the NuRD, Sin3A and Pml complexes, possibly to control ES fate. Reactivation of Oct-4 expression is postulated to occur in differentiated cells that have undergone carcinogenesis, or tumor formation.

Specificity: This product was affinity purified from monospecific antiserum by immunoaffinity chromatography. This antibody reacts with human and mouse Oct-4 protein. A BLAST analysis was used to suggest additional cross-reactivity with Oct-4 from chimpanzee, macaque and horse based on a 100% homology with the immunizing sequence, and mink, dog, bovine and pig based on a 93% homology. Cross-reactivity with Oct-4 from other sources has not been determined.

Application Details

Application Notes: Recommended Dilutions: ELISA 1:350,000 WESTERN BLOT 1:1,000 - 1:1,500 IMMUNOPRECIPITATION User Optimized IF MICROSCOPY User Optimized OTHER APPLICATIONS User Optimized. Note: This affinity purified antibody has been tested for use in ELISA and western blotting. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately ~39 kDa in size corresponding to Oct-4 by western blotting in the appropriate cell lysate or extract. This antibody is capable of detecting endogenous Oct-4..

Concentration: 1.0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20o C or below prior to opening. Dilute only prior to immediate use. For extended storage, aliquot contents and freeze at -20o C or below. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date of opening.

Restrictions: For Research Use only

Images

Western blot using affinity purified anti-Oct4 antibody shows detection of endogenous Oct4 in mouse embryonic fibroblast (MEF) cell lysate (lane 1) and HeLa nuclear extract (HN) (lane 2). The band at ~39 kDa (arrow head) corresponds to Oct4. After transfer, the membrane was blocked with 5% BSA. Primary antibody was used at a 1:1,000 dilution in PBS containing 5% BSA. The specificity of the antibody was confirmed by peptide competition which blocks reaction of the antibody with Oct4 (lanes 3 and 4, respectively). MEF HN MW MEF HN