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Sv40 t ag

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SV40 T Ag is a viral protein that is commonly used as a research tool in cell and molecular biology. It functions as a transcriptional regulator and can induce immortalization of cells.

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Lab products found in correlation

N cadherin, by Santa Cruz Biotechnology (2 mentions) Pan keratin, by Cell Signaling Technology (2 mentions) Nedd9, by Abcam (2 mentions) P130cas, by Santa Cruz Biotechnology (2 mentions) Psrcy527, by Cell Signaling Technology (2 mentions) P stat3 y705, by Cell Signaling Technology (2 mentions) Stat3, by Cell Signaling Technology (2 mentions) E cadherin, by Santa Cruz Biotechnology (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) Phospho s6 ribosomal protein ser240 244, by Cell Signaling Technology (1 mentions) Phospho s6 ribosomal protein ser235 236, by Cell Signaling Technology (1 mentions) Vectastain elite abc kit, by Vector Laboratories (1 mentions) Phospho akt s473, by Cell Signaling Technology (1 mentions) Anti erg antibody, by Abcam (1 mentions) H ras, by Santa Cruz Biotechnology (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Qiashredder, by Qiagen (1 mentions) Gapdh, by Merck Group (1 mentions) Anti actin, by Merck Group (1 mentions) Glucagon, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Anti-SV40 T Ag (6 citations) SV40 T-Ag antibody (2 citations)

8 protocols using sv40 t ag

1

Immunohistochemical Analysis of Mouse Stomach

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The stomachs taken from mice were opened in the minor curvature, fixed in 4% formaldehyde for 2 h at room temperature, dehydrated and embedded in paraffin wax. 2–3 μm sections were deparaffinized in xylene, and rehydrated in a graded series of ethanol. Antigen retrieval was performed when necessary in boiling natrium citrate buffer (PH6.0) for 15 min. Antibodies directed at Glucagon (Cell signaling,1:100), Chromogranin B (Santa Cruz, 1:100), SV40 T Ag (Santa Cruz, 1:500), Ki-67 (Dako, 1:50), Hydrogen Potassium ATPase Beta (Abcam, 1:1000), Phospho-S6 Ribosomal Protein (Ser235/236) (Cell signaling, 1:100), Phospho-S6 Ribosomal Protein (Ser240/244) (Cell signaling, 1:800), were applied to the sections for 2 h incubation at room temperature. Sections were then treated with HRP-coupled secondary antibodies (ImmPRESS Anti-Rabbit Ig Polymer Detection Kit, Vector Labs) or biotin-conjugated secondary antibody followed by avidin-biotin-peroxidase complex (VECTASTAIN Elite ABC Kit, Vector Labs). Primary antibodies were visualized by AEC or DAB as substrate.
Pan J., Bao Q, & Enders G. (2020). The Altered Metabolic Molecular Signatures Contribute to the RAD001 Resistance in Gastric Neuroendocrine Tumor. Frontiers in Oncology, 10, 546.
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2

Immunohistochemistry Staining Protocol

Cited 2 times
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Immunohistochemistry (IHC) was carried out using the general procedures described previously [49 (link)], using steam with Tris-EDTA buffer (ph 9.0) for antigen retrieval [14 (link)]. Detection was carried out at room temperature using a PolyVue Polymer detection system (Diagnostic Biosystems) according to manufacturer's directions. Primary antibodies were: anti-ERG antibody (#2805-1, Epitomics); phospho-AKT S473(#9271, Cell Signaling); androgen receptor (clone EPR 1535(2), Cat#3165-1, Epitomics); SV40 T Ag (Santa Cruz v-300, sc-20800); all at 1:100 dilution for 30 min at room temperature.
Shao L., Wang J., Karatas O.F., Feng S., Zhang Y., Creighton C.J, & Ittmann M. (2018). Fibroblast growth factor receptor signaling plays a key role in transformation induced by the TMPRSS2/ERG fusion gene and decreased PTEN. Oncotarget, 9(18), 14456-14471.
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3

Western Blot Analysis of Cellular Proteins

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Cells were lysed with 4% SDS and supernatant was collected after lysate centrifuged in Qiashredder (Qiagen). Protein concentrations were determined with BCA assay (Thermo Fisher Scientific: 23225). Equal amount of proteins (30 ug) were loaded into polyacrylamide SDS page gels. Candidate proteins were detected with antibodies against hTERT (Thermo Fisher Scientific: PA5-11447), SV40 T Ag (Santa Cruz Biotechnology: sc-147) and H-Ras (Santa Cruz Biotechnology: sc-520). GAPDH (Sigma: G9545) serves as a loading control.
Xie W., Kagiampakis I., Pan L., Zhang Y.W., Murphy L., Tao Y., Kong X., Xia L., Carvalho F.L., Sen S., Yen R.W., Zahnow C.A., Ahuja N., Baylin S.B, & Easwaran H. (2018). DNA methylation patterns separate senescence from transformation potential and indicate cancer risk. Cancer cell, 33(2), 309-321.e5.
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4

Western Blot Analysis of Viral and Cellular Proteins

Cited 1 time
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Western blotting was performed as described previously (17 (link)). The following primary antibodies were used for western blot: mouse mAb to HIRA (WC119 (30 (link))), actin (A1978; Sigma), MAb anti-HCMV-IE1 2470–5604 (AbD Serotec), HSV1-ICP8 MAb [11E2] ab20194 (Abcam), anti-UL42 MAb Z1F11 (31 (link)), anti-actin rabbit serum A5060 (Sigma-Aldrich), SV40 T Ag (Santa Cruz sc-147), p53 (Santa Cruz sc-126), pRb (Cell Signalling Technology 9309S).
Rai T.S., Glass M., Cole J.J., Rather M.I., Marsden M., Neilson M., Brock C., Humphreys I.R., Everett R.D, & Adams P.D. (2017). Histone chaperone HIRA deposits histone H3.3 onto foreign viral DNA and contributes to anti-viral intrinsic immunity. Nucleic Acids Research, 45(20), 11673-11683.
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5

Stomach Tissue Immunohistochemistry

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Tissue sections from the mice stomach were prepared as above. Antibodies directed at SV40 T Ag (Santa Cruz, 1:500), Glucagon (Cell signaling,1:100), Secretin (Biozol, 1:100) were used for primary incubation (2 h, room temperature). Sections were then reacted with appropriate secondary antibodies for 1 h at room temperature and counterstained with Hoechst33342.
Pan J., Bao Q, & Enders G. (2020). The Altered Metabolic Molecular Signatures Contribute to the RAD001 Resistance in Gastric Neuroendocrine Tumor. Frontiers in Oncology, 10, 546.
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6

Unmasking Tissue Antigens for Immunostaining

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Prior to immunostaining using the ImmunoCruz rabbit LSAB Staining System (Santa Cruz Biotechnology), tissue antigens were unmasked by heat treatment as per the manufacturer’s instructions. Briefly, prepared slides were treated with 10 mM sodium citrate buffer (pH 6) and heated to 95 °C. This process was then repeated with fresh buffer. After the slides cooled, they were washed with double distilled deionized H2O and the excess liquid was aspirated. After unmasking and blocking formalin-fixed 6 μm paraffin embedded tissues sections, antigens were detected using primary antibodies against luteinizing hormone receptor (LHR; Santa Cruz Biotechnology), AMH (Abcam, San Francisco, CA), SV40Tag (Santa Cruz Biotechnology), and mouse CD3 (Novacastra, Buffalo Grove, IL). All primary antibodies were used at a 1/1000 dilution.
Aguilar R., Johnson J.M., Barrett P, & Tuohy V.K. (2017). Vaccination with inhibin-α provides effective immunotherapy against testicular stromal cell tumors. Journal for Immunotherapy of Cancer, 5, 37.
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7

Derivation and Characterization of Ovarian Cancer Cell Lines

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MOVCAR cell lines were derived from the ascites of MISIIR-TAg;NEDD9+/+ (MOVCAR-5009, -6111, -7577, -8248, -8250) and MISIIR-TAg;NEDD9−/− (MOVCAR-136, -143, -145, -168) mice in our laboratory as described12 . Human OC cell lines OVCAR-5 and A1847 OC were obtained from the FCCC Cell Culture Facility (deposited by Dr. Thomas Hamilton) and maintained as described56 (link). Cell lines were periodically checked for mycoplasma contamination OVCAR-5 cells were authenticated by STR profiling. Antibodies and commercial source: antibodies recognizing AKT (cat# 468S), pAKTS473 (cat# 3787S), ERK 1/2 (9102), pERK1/2T202-Y204 (cat# 9106S), Src (cat# 2108S), pSrcY416 (cat# 2101S), pSrcY527 (2105S), STAT3 (cat# 9132) and pSTAT3Y705 (cat# 9145S), pan-keratin (cat# 4545) (Cell Signaling Technology; Danvers, MA, USA), NEDD9 (cat# ab18056) (Abcam; Cambridge, MA, USA), SV40-TAg (cat# sc-147), β-actin (cat# sc-8432), E-cadherin (cat# sc-7870), N-cadherin (cat# sc-7939), p130Cas (cat# sc-860) (Santa Cruz Biotechnology; Santa Cruz, CA, USA), BMPR1B (cat# bs-6639R), and FOXJ1 (cat# 36887) (One World Lab; San Diego, CA, USA), and PAX8 (cat# 10336-I-AP) (Proteintech Group, Rosemont, IL, USA).
Gabbasov R., Xiao F., Howe C.G., Bickel L.E., O’Brien S.W., Benrubi D., Do T.V., Zhou Y., Nicolas E., Cai K.Q., Litwin S., Seo S., Golemis E.A, & Connolly D.C. (2018). NEDD9 promotes oncogenic signaling, a stem/mesenchymal gene signature, and aggressive ovarian cancer growth in mice. Oncogene, 37(35), 4854-4870.
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8

Derivation and Characterization of Ovarian Cancer Cell Lines

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MOVCAR cell lines were derived from the ascites of MISIIR-TAg;NEDD9+/+ (MOVCAR-5009, -6111, -7577, -8248, -8250) and MISIIR-TAg;NEDD9−/− (MOVCAR-136, -143, -145, -168) mice in our laboratory as described12 . Human OC cell lines OVCAR-5 and A1847 OC were obtained from the FCCC Cell Culture Facility (deposited by Dr. Thomas Hamilton) and maintained as described56 (link). Cell lines were periodically checked for mycoplasma contamination OVCAR-5 cells were authenticated by STR profiling. Antibodies and commercial source: antibodies recognizing AKT (cat# 468S), pAKTS473 (cat# 3787S), ERK 1/2 (9102), pERK1/2T202-Y204 (cat# 9106S), Src (cat# 2108S), pSrcY416 (cat# 2101S), pSrcY527 (2105S), STAT3 (cat# 9132) and pSTAT3Y705 (cat# 9145S), pan-keratin (cat# 4545) (Cell Signaling Technology; Danvers, MA, USA), NEDD9 (cat# ab18056) (Abcam; Cambridge, MA, USA), SV40-TAg (cat# sc-147), β-actin (cat# sc-8432), E-cadherin (cat# sc-7870), N-cadherin (cat# sc-7939), p130Cas (cat# sc-860) (Santa Cruz Biotechnology; Santa Cruz, CA, USA), BMPR1B (cat# bs-6639R), and FOXJ1 (cat# 36887) (One World Lab; San Diego, CA, USA), and PAX8 (cat# 10336-I-AP) (Proteintech Group, Rosemont, IL, USA).
Gabbasov R., Xiao F., Howe C.G., Bickel L.E., O’Brien S.W., Benrubi D., Do T.V., Zhou Y., Nicolas E., Cai K.Q., Litwin S., Seo S., Golemis E.A, & Connolly D.C. (2018). NEDD9 promotes oncogenic signaling, a stem/mesenchymal gene signature, and aggressive ovarian cancer growth in mice. Oncogene, 37(35), 4854-4870.
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