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POU3F2 is a protein-coding gene that produces a transcription factor involved in the regulation of cell development and differentiation. The gene is expressed in various tissues and plays a role in the development of the nervous system and other cell types. This product can be used for research purposes to study the function and expression of the POU3F2 gene and protein.

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Lab products found in correlation

Gapdh, by Merck Group (2 mentions) Cell lysis buffer, by Beyotime (2 mentions) Detergent compatible bradford protein assay kit, by Beyotime (2 mentions) Beyoecl plus, by Beyotime (2 mentions) Trim8, by Abcam (2 mentions) P akt, by Cell Signaling Technology (1 mentions) Neurod1, by Abcam (1 mentions) Vinculin, by Cell Signaling Technology (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) P pras40, by Cell Signaling Technology (1 mentions) Ascl1, by BD (1 mentions) Aperio digital pathology slide scanner, by Leica (1 mentions) Imagescope software, by Leica (1 mentions) Mab1259, by R&D Systems (1 mentions) Rodent block m reagent, by Biocare Medical (1 mentions) Celltrace violet, by Thermo Fisher Scientific (1 mentions) Glial fibrillary acidic protein (gfap), by Merck Group (1 mentions) Lysotracker, by Thermo Fisher Scientific (1 mentions) Olig2, by Merck Group (1 mentions) Vinculin, by Merck Group (1 mentions)

5 protocols using pou3f2

1

Protein Expression Analysis by Western Blot

Cited 1 time
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Protein extraction and western blot were performed as previously described(68 (link)). Antibodies for FOXN4 (#PA539174, ThermoFisher), ONECUT2 (#ab28466, Abcam), POU3F2 (#12137, Cell Signaling Technology), EGFR (#4267, Cell Signaling Technology), ASCL1 (#556604, BD), NEUROD1 (#ab109224, Abcam), pAKT (S473, #4060S, Cell Signaling Technology), pPRAS40 (T246, #13175, Cell Signaling Technology), Beta catenin (#8480, Cell Signaling Technology), Vinculin (#13901, Cell Signaling Technology) and actin (#3700, Cell Signaling Technology) were used.
Quintanal-Villalonga A., Taniguchi H., Zhan Y.A., Hasan M.M., Chavan S.S., Meng F., Uddin F., Manoj P., Donoghue M.T., Won H.H., Chan J.M., Ciampricotti M., Chow A., Offin M., Chang J.C., Ray-Kirton J., Tischfield S.E., Egger J., Bhanot U.K., Linkov I., Asher M., Sinha S., Silber J., Iacobuzio-Donahue C.A., Roehrl M.H., Hollmann T.J., Yu H.A., Qiu J., de Stanchina E., Baine M.K., Rekhtman N., Poirier J.T., Loomis B., Koche R.P., Rudin C.M, & Sen T. (2021). Multi-omic analysis of lung tumors defines pathways activated in neuroendocrine transformation. Cancer discovery, 11(12), 3028-3047.
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2

Immunohistochemical Analysis of Glioblastoma Samples

Cited 2 times
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Brains from mice inoculated with vehicle (controls) or primary GBM NS characterized by low or high V-ATPase G1 expression were harvested at sacrifice and formalin-fixed and paraffin embedded (FFPE) as described in the companion study (Terrasi et al., this issue) [36 ]. Then, hematoxylin and eosin (H&E) staining was performed to confirm tissue morphology followed by immunohistochemistry for HOXA10 (TA590263, Origen), STEM121 (Y40410, Takara Bio Europe) and POU3F2 (12137, Cell Signaling). Human tissues were already FFPE blocks. IHC for V-ATPase G1 (16143-1-AP, Proteintech), Nestin (MAB1259, R&D Systems), HOXA10, or POU3F2 was performed using a Ventana Benchmark instrument and the Ultraview DAB or Red Detection kits as described [13 (link)]. For murine samples, slides were pre-incubated with the Rodent Block M reagent (Biocare Medical), to block endogenous mouse IgG and non-specific background, before primary antibodies. Percentage of positive tumor cells in PDTX tumors was analyzed using Aperio Digital Pathology slide scanner (Leica Biosystems, Milan, Italy) and the nuclear algorithm implemented in ImageScope software (Leica Biosystems) as described [15 (link)].
Bertolini I., Terrasi A., Martelli C., Gaudioso G., Di Cristofori A., Storaci A.M., Formica M., Braidotti P., Todoerti K., Ferrero S., Caroli M., Ottobrini L., Vaccari T, & Vaira V. (2019). A GBM-like V-ATPase signature directs cell-cell tumor signaling and reprogramming via large oncosomes. EBioMedicine, 41, 225-235.
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3

Multiparametric Analysis of Cell Cultures

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Lysotracker, CellTrace Violet, FM lipophilic styryl (FM 1-43FX) and SYTO RNASelect dyes were from Life Technologies (Thermo Fisher Scientific). Acridine Orange was from ImmunoChemistry Technologies (Bloomington, MN, USA). PropidiumIodure (PI) and Annexin V were from BD Bioscience (FITC-Annexin V Apoptosis Detection Kit). Bafilomycin A1 (BafA1; sc-201550) was from Santa Cruz Biotechnologies whereas NH4Cl (254134) was from Sigma Aldrich.
The following primary antibodies were used for immunofluorescence, immunoblotting (IB) and flow cytometry (FACS) assays: V-ATPase G1 (16143-1-AP, Proteintech), Vinculin (V9131, Sigma Aldrich), Tsg101 (14497-1-AP, Proteintech), Ago2 (10686-1-AP, Proteintech), Clathrin (ab23440, Abcam), CD63 (sc-15363, Santa Cruz), CD9 (IB: 10626D, Thermo Fisher; FACS: 130-103-988, Miltenyi), CD81 (130-107-982, Miltenyi), Calnexin (ab31290, Abcam), Nestin (MAB 1259, R&D), Tuji (T3952, Sigma Aldrich), GFAP (G9269, Sigma Aldrich), CD11b (20991-1-AP, Proteintech), O4 (O7139, Sigma Aldrich), Olig2 (AV32753, Sigma Aldrich), CD31 (ab28364, Abcam), Vimentin (130-106-369, Miltenyi), POU3F2 (12137, Cell Signaling), HOXA10 (TA590263, Origen), HOXA7 (ab211521, Abcam), V-ATPase G2 (25316-1-AP, Proteintech), ARF6 (6ARF01, ThermoFisher Scientific), and GRP78 (3177, Cell Signaling).
Bertolini I., Terrasi A., Martelli C., Gaudioso G., Di Cristofori A., Storaci A.M., Formica M., Braidotti P., Todoerti K., Ferrero S., Caroli M., Ottobrini L., Vaccari T, & Vaira V. (2019). A GBM-like V-ATPase signature directs cell-cell tumor signaling and reprogramming via large oncosomes. EBioMedicine, 41, 225-235.
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4

Protein Isolation and Western Blot Analysis

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Protein isolation was performed with cell lysis buffer (Beyotime, #P0013). Protein concentration was detected using a Detergent Compatible Bradford Protein Assay Kit (Beyotime). Proteins were separated by SDS-polyacrylamide gel electrophoresis, transferred to PVDF membranes, and blocked by 5% milk for 1.5 h. Membranes were incubated with primary antibodies overnight at 4°C. The following antibodies were used: POU3F2 (1:1000, Cell Signaling Technology 12137), TRIM8 (1:1000, Abcam ab155674), and GAPDH (1:10000, Sigma G9295). Corresponding secondary antibodies (1:5000, Jackson ImmunoResearch 111–035-144) were used to bind with primary antibodies at room temperature for 2 h. Protein bands were detected by chemiluminescence using the BeyoECL Plus (Beyotime, #P0018S). Relative intensities of the protein bands were quantified by software ImageJ (https://imagej.nih.gov/ij/).
Ding C., Zhang C., Kopp R., Kuney L., Meng Q., Wang L., Xia Y., Jiang Y., Dai R., Min S., Yao W.D., Wong M.L., Ruan H., Liu C, & Chen C. (2020). Transcription factor POU3F2 regulates TRIM8 expression contributing to cellular functions implicated in schizophrenia. Molecular psychiatry, 26(7), 3444-3460.
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5

Protein Isolation and Western Blot Analysis

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Protein isolation was performed with cell lysis buffer (Beyotime, #P0013). Protein concentration was detected using a Detergent Compatible Bradford Protein Assay Kit (Beyotime). Proteins were separated by SDS-polyacrylamide gel electrophoresis, transferred to PVDF membranes, and blocked by 5% milk for 1.5 h. Membranes were incubated with primary antibodies overnight at 4°C. The following antibodies were used: POU3F2 (1:1000, Cell Signaling Technology 12137), TRIM8 (1:1000, Abcam ab155674), and GAPDH (1:10000, Sigma G9295). Corresponding secondary antibodies (1:5000, Jackson ImmunoResearch 111–035-144) were used to bind with primary antibodies at room temperature for 2 h. Protein bands were detected by chemiluminescence using the BeyoECL Plus (Beyotime, #P0018S). Relative intensities of the protein bands were quantified by software ImageJ (https://imagej.nih.gov/ij/).
Ding C., Zhang C., Kopp R., Kuney L., Meng Q., Wang L., Xia Y., Jiang Y., Dai R., Min S., Yao W.D., Wong M.L., Ruan H., Liu C, & Chen C. (2020). Transcription factor POU3F2 regulates TRIM8 expression contributing to cellular functions implicated in schizophrenia. Molecular psychiatry, 26(7), 3444-3460.
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