Anti prps1

Manufactured by Proteintech

Sourced in United States

Anti-PRPS1 is a lab equipment product that is used to detect and measure the expression levels of the PRPS1 protein. PRPS1 is an enzyme involved in the production of purine nucleotides, which are essential components of DNA and RNA. The Anti-PRPS1 product provides researchers with a tool to study the role of PRPS1 in various biological processes and disease states.

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3 protocols using anti prps1

Western Blot Profiling of Protein Expression

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The cells were prepared in RIPA buffer (Solarbio, #R0020). A BCA™ Protein Assay kit (Applygen, #P1511) was used to determine the protein concentration. The proteins (40 μg/sample) were separated by different polyacrylamide gel electrophoresis, transferred to PVDF membranes (Millipore, #IPVH00010), and incubated with the corresponding primary antibody at 4°C overnight. Then, the membranes were incubated with the corresponding secondary antibodies at room temperature for 1 h and measured with a chemiluminescence reagent ECL kit (Advansia, #K-12045-D50).
The primary antibodies used in the experiment used were: anti-PRPS1 (Proteintech, #15549-1-AP), anti-cyclin E1 (Proteintech, 11554-1-AP), anti-CDK2 (Proteintech, 10122-1-AP), anti-P16 (Proteintech, #10883-1-AP), anti-Bax (Proteintech, 50599-2-Ig), anti-Bcl2 (Proteintech, 12789-1-AP), anti-Cleaved-caspeas3 (CST, #9664), anti-MMP2 (Abcam, ab37150), anti-MMP9 (Abcam, ab76003), anti-MMP13 (Proteintech, 18165-1-AP), anti-E-Cadherin (Proteintech, 20874-1-AP), anti-N-Cadherin (Proteintech, 22018-1-AP), anti-Vimentin (Proteintech, 10366-1-AP), anti-NRF2 (Abcam, ab89443), anti-β-actin (Bioss, bs-0061R), and Tubulin (Abcam, #ab7291). The secondary antibodies used in the experiment were anti-rabbit IgG (Abcam, #ab6721) and anti-mouse IgG (Jackson ImmunoResearch Laboratories, 115-035-003).

Xiong G., Feng Y., Yi X., Zhang X., Li X., Yang L., Yi Z., Sai B., Yang Z., Zhang Q., Kuang Y, & Zhu Y. (2022). NRF2-directed PRPS1 upregulation to promote the progression and metastasis of melanoma. Frontiers in Immunology, 13, 989263.

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Western Blot Analysis of Protein Expression

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Cells were solubilized in lysis buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% Nonidet P-40, 5 mM EDTA, 0.05% NaN3, 1 mM phenylmethylsulfonyl fluoride, 100 µM sodium vanadate) on ice. The cell lysates were separated on a SDS-polyacrylamide gel under reducing conditions and transferred to a nitrocellulose membrane.
Each membrane was incubated with anti-NT5C2 (Abcam, MA, USA, ab96084), anti-PRPS1 (Proteintech, Tokyo, Japan, 15549-1-AP), anti-beta-actin (MBL, PM053-7), and anti-alpha-tubulin (Sigma-Aldrich, T5168) antibodies at 4°C overnight. Subsequently, the membranes were incubated with horseradish peroxidase-labeled anti-mouse or anti-rabbit IgGs (MBL) at room temperature for 1 hour. Then, the blots were developed using an enhanced chemiluminescence detection (ECL) kit (GE Healthcare, Little Chalfont, UK). The band density was semi-quantified using Image J (National Institute of Health, MD, USA).

Nguyen T.T.T., Tanaka Y., Sanada M., Hosaka M., Tamai M., Kagami K., Komatsu C., Somazu S., Harama D., Kasai S., Watanabe A., Akahane K., Goi K, & Inukai T. (2023). CRISPR/Cas9-Mediated Induction of Relapse-Specific NT5C2 and PRPS1 Mutations Confers Thiopurine Resistance as a Relapsed Lymphoid Leukemia Model. Molecular pharmacology, 103(4).

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Glycolytic Enzyme Profiling of Tissue

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Specimens were excised by standard surgical procedures and were cut into 4 mm sections. Serial sections were prepared and selected for staining with H&E (Baso, Taipei, Taiwan). Before staining, the sections were incubated with 0.1% BSA, then incubated overnight at 4 C with a target protein antibody. Biotinylated IgG was used as a secondary antibody, followed by streptavidin-conjugated horseradish peroxidase.
Target proteins were analyzed using the following antibodies: anti-GYS1, anti-PYGL, anti-CA9, anti-phosphogluconate dehydrogenase, anti-PRPS1, anti-HK2, anti-PFKP, anti-pyruvate dehydrogenase beta, anti-citrate synthase, anti-isocitrate dehydrogenase 3A, and anti-oxoglutarate dehydrogenase were purchased from Proteintech (Wuhan, China), anti-lactate dehydrogenase A and anti-HIF-1a were purchased from Goodbio Company (Wuhan, China). The species source of all the antibody was rabbit.
Sample sections were stained with periodic acid-Schiff (Baso) sets to detect glycogen levels according to the manufacturer's instructions.

Gu Z., Zhang H., Guo X, & Cao Y. (2020). Enhanced Glycogen Metabolism Supports the Survival and Proliferation of HPV-Infected Keratinocytes in Condylomata Acuminata. The Journal of investigative dermatology, 140(8).

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