53-5841-80

antibody from Invitrogen Antibodies

Targeting: POU5F1 MGC22487, OCT3, Oct4, OTF3

Provider product page for 53-5841-80

Immunocytochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [2]
Comments [0]
Validations
Immunocytochemistry [1]
Flow cytometry [2]
Other assay [2]

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Product number: 53-5841-80 - Provider product page
Provider: Invitrogen Antibodies
Product name: Anti-OCT3/4 Monoclonal Antibody (EM92), Alexa Fluor 488, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: 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. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human, Mouse
Host: Rat
Conjugate: Green dye
Isotype: IgG
Antibody clone number: EM92
Vial size: 25 µg
Concentration: 0.5 mg/mL
Storage: 4° C, store in dark, DO NOT FREEZE!

POU5F1 protein structure - 53-5841-80 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of OCT3/4 was performed using 70% confluent log phase NTERA-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with OCT3/4 Mouse Monoclonal Antibody (Product # 53-5841-80) at 5ug/ml in 0.1% BSA, incubated at 4 degree Celsius overnight (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows OCT3/4 negative cell line HeLa with no signal. Panel f represents control cells with Isotype control to assess background. The images were captured at 60X magnification.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of OCT3/4 was performed using 70% confluent log phase NTERA cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with OCT3/4 Mouse Monoclonal Antibody (Product # 53-5841-82) at 5ug/ml in 0.1% BSA, incubated at 4 degree Celsius overnight (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows OCT3/4 negative cell line HeLa with no signal. Panel f represents control cells with Isotype control to assess background. The images were captured at 60X magnification.
Conjugate: Green dye