|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rat / IgG2a, kappa|
|Conjugate||Alexa Fluor® 488|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1 µg/test|
|Immunocytochemistry (ICC)||10 µg/ml|
|Immunofluorescence (IF)||10 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Description: The EM92 monoclonal antibody reacts with mouse and human Oct3/4, encoded by the Pou5F1 gene. Oct3/4 is a POU domain-containing transcription factor that is critical for maintaining embryonic stem (ES) and induced pluripotent stem (iPS) cells in a pluripotent state, and is expressed by ES, embryonic germ cells and embryonic carcinoma cell lines. In cells of the inner cell mass (ICM), reduction of Oct3/4 expression causes dedifferentiation to trophoectoderm, whereas increased expression results in differentiation to mesoderm and primitive endoderm. Oct3/4 regulates the expression of several genes, including FGF-4, UTF1, Sox2, Fbx15, Rex1 and osteopontin through distinct mechanisms. Furthermore, Oct3/4 frequently acts synergistically with Sox2 to regulate target gene expression, as is the case with FGF-4. It has been demonstrated that Oct3/4 expression in ES cells can be negatively regulated by either treatment with retinoic acid, or by removal of leukemia-inhibitory factor (LIF).
Applications Reported: This EM92 antibody has been reported for use in intracellular staining followed by flow cytometric analysis and immunocytochemistry.
Applications Tested: This EM92 antibody has been tested by microscopy or intracellular staining and flow cytometric analysis of F9 cells using the Foxp3/Transcription Factor Staining Buffer Set (cat. 00-5523) and protocol. Please see Best Protocols for Staining Protocol (refer to Protocol B: One step protocol for intracellular (nuclear) proteins). This antibody can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. This antibody has also been tested by immunocytochemistry on formaldehyde fixed and permeabilized cells at less than or equal to 10µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Excitation: 488 nm; Emission: 519 nm; Laser: Blue Laser
POU5F1, also commonly known as Oct-4, is a maternally expressed octamer-binding protein that was the first transcription factor described for the early stages of development. The role of POU5F1 in embryonic development suggested that it might be useful in the creation of stem cells that might be useful in cell replacement therapies in the treatment of several degenerative diseases. Artificial stem cells, termed induced pluripotent stem (iPS) cells, can be created by expressing POU5F1 and the transcription factors Sox2, Klf4 and Lin28 along with c-Myc in mouse fibroblasts. More recently, experiments have demonstrated that iPS cells could be generated using expression plasmids expressing POU5F1, Sox2, KlfF4 and c-Myc, eliminating the need for virus introduction, thereby addressing a safety concern for potential use of iPS cells in regenerative medicine.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.