Ptch1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

PTCH1 is a gene that encodes the Patched 1 protein. Patched 1 is a receptor that plays a key role in the Hedgehog signaling pathway, which is important for embryonic development and cell differentiation. The core function of PTCH1 is to act as a negative regulator of this signaling pathway.

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Lab products found in correlation

Ripa buffer, by Thermo Fisher Scientific (4 mentions) Gapdh, by Santa Cruz Biotechnology (2 mentions) Fancd2, by Santa Cruz Biotechnology (2 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Htra2, by Santa Cruz Biotechnology (1 mentions) Amersham ecl prime detection reagent, by GE Healthcare (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) γh2ax, by Merck Group (1 mentions) Laemmli buffer, by Bio-Rad (1 mentions) Phosphatase inhibitor, by Roche (1 mentions) Axiocam camera, by Zeiss (1 mentions) Axioscope microscope, by Zeiss (1 mentions) Igfbp 2, by Santa Cruz Biotechnology (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions) Supersignal west pico, by Thermo Fisher Scientific (1 mentions) β actin, by Abcam (1 mentions) Hif 1α, by Abcam (1 mentions)

5 protocols using ptch1

Immunoblotting Protein Analysis Protocol

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For the immunoblotting, a small amount (10 μL) of the CPB solution (0.5 McF), was mixed with Laemmli Buffer at a ratio 4:1, and loaded on 1.0 mm NuPAGE® Novex® 4–12% Bis-Tris protein gradient gel. Normal human liver protein extract was used as a positive control. Proteins were separated by the SDS-PAGE at 100 V for 1 h and transferred at 30 V for 1.5 h 4 °C. Blots were probed with either fetuin-A (ab34505, Abcam) or albumin (ab9092, Abcam) antibodies. Horseradish peroxidase conjugated rabbit or goat secondary antibodies (Santa Cruz Biotechnology) were used at dilution 1:500. Proteins were visualised with enhanced chemiluminescence by using the Amersham ECL Prime detection reagent (General Electric Healthcare). For the experiments described in Figs 6e and 7a,b, total protein was extracted from cells using RIPA buffer (ThermoFisher) with protease inhibitor cocktail (Roche). Electrophoretically resolved protein samples were transferred and probed with cleaved caspase 9 (GTX86912, GenTex), xiap (sc-11426, Santa Cruz Biotechnology), htra2 (sc-15467, Santa Cruz Biotechnology), shh (sc-9024, Santa Cruz Biotechnology), ptch1 (sc-9016, Santa Cruz Biotechnology), and gapdh (sc-20357, Santa Cruz Biotechnology) antibodies.

Kutikhin A.G., Velikanova E.A., Mukhamadiyarov R.A., Glushkova T.V., Borisov V.V., Matveeva V.G., Antonova L.V., Filip’ev D.E., Golovkin A.S., Shishkova D.K., Burago A.Y., Frolov A.V., Dolgov V.Y., Efimova O.S., Popova A.N., Malysheva V.Y., Vladimirov A.A., Sozinov S.A., Ismagilov Z.R., Russakov D.M., Lomzov A.A., Pyshnyi D.V., Gutakovsky A.K., Zhivodkov Y.A., Demidov E.A., Peltek S.E., Dolganyuk V.F., Babich O.O., Grigoriev E.V., Brusina E.B., Barbarash O.L, & Yuzhalin A.E. (2016). Apoptosis-mediated endothelial toxicity but not direct calcification or functional changes in anti-calcification proteins defines pathogenic effects of calcium phosphate bions. Scientific Reports, 6, 27255.

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Western Blotting for Protein Analysis

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For protein analysis by western blotting, cells were washed with ice-cold PBS and lysed on ice using ice-cold cytoskeletal (CSK) buffer [10 mM PIPES (pH 6.8), 100 mM NaCl, 300 mM sucrose, 3 mM MgCl₂, 1 mM EGTA, 1 mM dithiothreitol, 0.1 mM ATP, 1 mM Na₃VO₄, 10 mM NaF and 0.1% Triton X-100], freshly supplemented with protease (Cat No: 05892970001, Roche) and phosphatase inhibitors (Cat No: 04906837001, Roche). Lysed samples were normalized and diluted with 2X laemmli buffer (Cat No: 161-0737, Bio-Rad) and heated to 100°C for 10 minutes. Then the denatured samples were resolved by SDS-PAGE, transferred to nitrocellulose membranes and probed with the specific antibodies such as γH2AX (Cat No: 05636, Millipore), FANCD2 (Cat No: 20022, Santa Cruz), GLI1 (Cat No: 2553, Cell Signaling), PTCH1 (Cat No: 6147, Santa Cruz) and GAPDH (Cat No: 32233, Santa Cruz).

Mani C., Tripathi K., Chaudhary S., Somasagara R.R., Rocconi R.P., Crasto C., Reedy M., Athar M, & Palle K. (2021). Hedgehog/GLI1 Transcriptionally Regulates FANCD2 in Ovarian Tumor Cells: Its Inhibition Induces HR-Deficiency and Synergistic Lethality with PARP Inhibition.. Neoplasia (New York, N.Y.), 23(9), 1002-1015.

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Immunohistochemical Analysis of FANCD2, PTCH1, and GLI1

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Normal and mice tumor tissues were stained for the expression of FANCD2, PTCH1 and GLI1 proteins by immunohistochemistry. Tissue sections were incubated with specific antibodies FANCD2 (Cat No: 20022, Santa Cruz), PTCH1 (Cat No: 6147, Santa Cruz) and GLI1 (Cat No: 20687, Santa Cruz) followed by a specific biotinylated secondary antibody (1:250 dilution), and then conjugated HRP streptavidin and DAB chromogen, and tissues were counterstained with hematoxylin. Stained sections were analyzed by Zeiss Axioscope microscope and as described previously [26 (link)] images were captured by AxioCam camera.

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Gemcitabine and Signaling Pathway Evaluation

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Gemcitabine was purchased from Eli Lily (France). The following antibodies were used in this study: IGFBP-2, PTCH-1and Gli1 (Santa Cruz Biotechnology, CA, USA); E-cadherin, N-cadherin and vimentin (Cell Signaling Technology, Inc., MA, USA); and Ki-67 (Abcam Inc., MA, USA).

Liu H., Li L., Chen H., Kong R., Pan S., Hu J., Wang Y., Li Y, & Sun B. (2017). Silencing IGFBP-2 decreases pancreatic cancer metastasis and enhances chemotherapeutic sensitivity. Oncotarget, 8(37), 61674-61686.

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Western Blot Analysis of Hedgehog Pathway Proteins

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After the indicated treatment, cells were lysed with RIPA buffer (Thermo, Rockford, IL) mixed with protease inhibitor cocktail (Thermo, Rockford, IL). Protein concentrations were determined using Pierce BCA Protein Assay Kit (Thermo, Rockford, IL). Equal amounts of protein samples (30 μg/well) were separated electrophoretically by SDS-PAGE, and transferred to PVDF membranes (Roche, Mannheim, Germany). The membranes were blocked for 1 hour in PBS-Tween 20 with 5% bovine serum albumin. Thereafter, the blots were probed with primary antibodies against HIF-1α (1:1000), GLI1 (1:500), β-actin (1:1000; all from Abcam, Cambridge, MA), SHH (1:1000; Cell Signaling Technology, Danvers, MA), and PTCH1 (1:500; Santa Cruz Biotechnology, Santa Cruz, CA). The blots were washed in PBS-Tween 20, incubated with the appropriate secondary antibodies (horseradish peroxidase conjugated), and visualized using Super Signal West Pico (Thermo, Rockford, IL).

Chen S., Zhang M., Xing L., Wang Y., Xiao Y, & Wu Y. (2015). HIF-1α Contributes to Proliferation and Invasiveness of Neuroblastoma Cells via SHH Signaling. PLoS ONE, 10(3), e0121115.

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