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Ab170952

Manufactured by Abcam
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Sourced in United Kingdom

Ab170952 is a primary antibody product manufactured by Abcam. It is designed for use in various laboratory applications, including immunoassays and immunohistochemistry. The product details and technical specifications are available on the Abcam website.

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Lab products found in correlation

Pvdf membrane, by Merck Group (2 mentions) A1978, by Merck Group (2 mentions) A2856, by Merck Group (2 mentions) Sc 374353, by Santa Cruz Biotechnology (2 mentions) Sc 515791, by Santa Cruz Biotechnology (2 mentions) Sc 22597, by Santa Cruz Biotechnology (2 mentions) Ab108327, by Abcam (2 mentions) Chemidoc xrs system, by Bio-Rad (2 mentions) Sc 271430, by Santa Cruz Biotechnology (2 mentions) Nel105001ea, by PerkinElmer (2 mentions) Ab40800, by Abcam (2 mentions) S1699, by Agilent Technologies (1 mentions) Roti histofix, by Carl Roth (1 mentions) Bca protein quantification kit, by Beyotime (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions) Ab246514, by Abcam (1 mentions) Gapdh 5174, by Cell Signaling Technology (1 mentions) Ab51520, by Abcam (1 mentions) Ab203859, by Abcam (1 mentions) Ab194586, by Abcam (1 mentions)

8 protocols using ab170952

1

Immunohistochemical Profiling of Liver Tissue

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Liver pieces were fixed in Roti Histofix (P087.4, Carl Roth) for 48h. Afterward the fixation solution was exchanged to 70% (v/v) ethanol and stored until paraffin-embedment. For immunohistochemical stainings 2 μm FFPE consecutive sections cut, heat-mediated antigen retrieval was performed in citrate buffer at pH 6.0 (S1699; Dako, Agilent, Santa Clara, CA, USA). Sections were stained with antibodies specific to STAT1 monoclonal rabbit (1:200; 9172; Cell Signaling Technology), ASS1 monoclonal rabbit (1:400; ab170952; Abcam), OTC monoclonal rabbit (1:500, sc-5157791; Santa Cruz), using standard protocols.
Lercher A., Bhattacharya A., Popa A.M., Caldera M., Schlapansky M.F., Baazim H., Agerer B., Gürtl B., Kosack L., Májek P., Brunner J.S., Vitko D., Pinter T., Genger J.W., Orlova A., Pikor N., Reil D., Ozsvár-Kozma M., Kalinke U., Ludewig B., Moriggl R., Bennett K.L., Menche J., Cheng P.N., Schabbauer G., Trauner M., Klavins K, & Bergthaler A. (2019). Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function. Immunity, 51(6), 1074-1087.e9.
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2

Western Blotting Protein Analysis Protocol

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The Western blotting assay was conducted in accordance with a previous study [24 (link)]. The cell lysate containing the protease inhibitor, phenylmethylsulfonyl fluoride (PMSF) (Beyotime, Nantong, China), was added to the cells to extract proteins and quantified using a BCA protein quantification kit (Beyotime, Nantong, China). Equal amounts of proteins were added to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the membrane was transferred, and the polyvinylidene fluoride (PVDF) membrane of the corresponding size was cut out according to the molecular weight. After blocking with 5% skimmed milk in Tris-buffered saline with Tween 20 (TBST) for 2 h, the membranes were incubated with the corresponding primary antibodies (ASS1, 1/2000, ab170952, Abcam; YTHDF2, 1/1000, ab246514, Abcam) at 4 °C overnight. The membranes were then incubated with horseradish peroxidase-conjugated secondary antibodies for 2 h at room temperature. Immunoblots were detected using an imaging system (Bio-Rad, USA). GAPDH was used as the loading control.

Miao Y.Q., Chen W., Zhou J., Shen Q., Sun Y., Li T, & Wang S.C. (2022). N(6)-adenosine-methyltransferase-14 promotes glioma tumorigenesis by repressing argininosuccinate synthase 1 expression in an m6A-dependent manner. Bioengineered, 13(1), 1858-1871.
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3

Antibody Immunoblotting Protocol

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Antibodies against ASS1 (ab170952), ASL (ab201026), Beclin 1 (ab210498), LC3B (ab51520), OTC (ab203859) and PARP1 (ab194586) were obtained from abcam (Cambridge, UK). Antibodies against β-ACTIN (MA515730) were obtained Thermo Fisher Scientific. Antibodies against AMPKα (2532), Phospho-AMPKα (Thr172) (2535S) and GAPDH (5174S) were obtained from Cell Signaling (Danvers, MA). Immunoblotting was performed as previously described [18 (link)].
Chow J.P., Cai Y., Chow D.T., Chung S.H., Chau K.C., Ng K.Y., Leung O.M., Wong R.M., Law A.W., Leung Y.O., Kwok S.Y, & Leung Y.C. (2020). A modified arginine-depleting enzyme NEI-01 inhibits growth of pancreatic cancer cells. PLoS ONE, 15(4), e0231633.
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4

Western Blot Analysis of Urea Cycle Enzymes

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Tissues and tumor samples were grounded in liquid nitrogen, lysed in Tris lysis buffer (50 mM Tris HCl, 150 mM NaCl, 1 mM EDTA, 0.1% NP40, 5 mM MgCl2, 10% glycerol), separated on 12.5% SDS-PAGE gel and then transferred on PVDF membrane (Millipore). Membranes were blocked with 5% non-fat milk for 1 hour and probed overnight at 4˚C with antibodies against ASS1 (1:1,000, Ab170952, Abcam), ASL (1:500, sc-374353, Santa Cruz), OTC (1:500, sc-515791, Santa Cruz), ARG1 (1:500, sc-271430, Santa Cruz), ATG7 (1:2,000, A2856, Sigma), transferrin (1:1,000, sc-22597, Santa Cruz), ATG5 (1:1,500, Ab108327, Abcam) and β-actin (1:5,000, A1978, Sigma). Immunoreactive bands were detected using peroxidase-conjugated antibody (GE Healthcare) and enhanced chemiluminescence detection reagents (NEL105001EA, Perkin Elmer) and were analyzed using the ChemiDoc XRS+ system (Biorad). Protein levels were quantified using the Image Lab v6.0.1 software. Antibodies were validated with the use of positive and negative control following manufacturer’s protocol.
Poillet-Perez L., Xie X., Zhan L., Yang Y., Sharp D.W., Hu Z.S., Su X., Maganti A., Jiang C., Lu W., Zheng H., Bosenberg M.W., Mehnert J.M., Guo J.Y., Lattime E., Rabinowitz J.D, & White E. (2018). Autophagy maintains tumor growth through circulating arginine. Nature, 563(7732), 569-573.
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5

Immunohistochemical Analysis of Arginine Metabolism

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Paraffin-embedded tissue sections (4 μm) were soaked in xylene for three times, then rehydration through an ethanol to water gradient at 100%/100%/95%/ 90%/80%/70%. Then, sections were placed in Citrate Antigen Retrieval Solution (0.1 M, pH 6.0) and heated for 5 min. After cooling to room temperature (RT), sections were blocked with 5% BSA for 1 h, then incubated with the anti-arginase antibody (ab233548, EPR22033-369, abcam, England, 1:1000 dilution) and anti-ASS1 antibody (ab170952, EPR12398, abcam, 1:1000 dilution) at 4 °C overnight, or incubated with the anti-iNOS antibody (ab283655, RM1017, abcam, 1:200 dilution) and anti-F4/80 antibody (ab300421, EPR26545-166, abcam, 1:10,000 dilution) at 37 °C for 1 h. After that, sections were washed five times and then incubated with the appropriate secondary antibody (PV-6001, ZSGB-BIO, Beijing, China) for 20 min at RT. Finally, the tissue sections were counterstained with DAPI (C1005, Beyotime) for nuclei visualization. Images were acquired using a scanister (Tissue Gnostics, Austria). To allow comparison between groups, fluorescence intensity was measured using ImageJ software.
Yang H., Wu X., Li X., Zang W., Zhou Z., Zhou Y., Cui W., Kou Y., Wang L., Hu A., Wu L., Yin Z., Chen Q., Chen Y., Huang Z., Wang Y, & Gu B. (2024). A commensal protozoan attenuates Clostridioides difficile pathogenesis in mice via arginine-ornithine metabolism and host intestinal immune response. Nature Communications, 15, 2842.
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6

Protein Expression Quantification Assay

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The assay was performed as published21 (link) using Sigma Aldrich kit (Cat # DUO 92004-30-RXN). Antibodies used for detection were diluted in PBS; ASS1 (1:200, #ab170952, abcam), citrin (1:100, #H00010165-M01, clone # 4F4, abnova) and anti-CAD (1:100, ab40800, abcam).
Rabinovich S., Adler L., Yizhak K., Sarver A., Silberman A., Agron S., Stettner N., Sun Q., Brandis A., Helbling D., Korman S., Itzkovitz S., Dimmock D., Ulitsky I., Nagamani S.C., Ruppin E, & Erez A. (2015). Diversion of aspartate in ASS1-deficient tumors fosters de novo pyrimidine synthesis. Nature, 527(7578), 379-383.
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7

Western Blot Analysis of Urea Cycle Enzymes

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Tissues and tumor samples were grounded in liquid nitrogen, lysed in Tris lysis buffer (50 mM Tris HCl, 150 mM NaCl, 1 mM EDTA, 0.1% NP40, 5 mM MgCl2, 10% glycerol), separated on 12.5% SDS-PAGE gel and then transferred on PVDF membrane (Millipore). Membranes were blocked with 5% non-fat milk for 1 hour and probed overnight at 4˚C with antibodies against ASS1 (1:1,000, Ab170952, Abcam), ASL (1:500, sc-374353, Santa Cruz), OTC (1:500, sc-515791, Santa Cruz), ARG1 (1:500, sc-271430, Santa Cruz), ATG7 (1:2,000, A2856, Sigma), transferrin (1:1,000, sc-22597, Santa Cruz), ATG5 (1:1,500, Ab108327, Abcam) and β-actin (1:5,000, A1978, Sigma). Immunoreactive bands were detected using peroxidase-conjugated antibody (GE Healthcare) and enhanced chemiluminescence detection reagents (NEL105001EA, Perkin Elmer) and were analyzed using the ChemiDoc XRS+ system (Biorad). Protein levels were quantified using the Image Lab v6.0.1 software. Antibodies were validated with the use of positive and negative control following manufacturer’s protocol.
Poillet-Perez L., Xie X., Zhan L., Yang Y., Sharp D.W., Hu Z.S., Su X., Maganti A., Jiang C., Lu W., Zheng H., Bosenberg M.W., Mehnert J.M., Guo J.Y., Lattime E., Rabinowitz J.D, & White E. (2018). Autophagy maintains tumor growth through circulating arginine. Nature, 563(7732), 569-573.
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8

Protein Expression Quantification Assay

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The assay was performed as published21 (link) using Sigma Aldrich kit (Cat # DUO 92004-30-RXN). Antibodies used for detection were diluted in PBS; ASS1 (1:200, #ab170952, abcam), citrin (1:100, #H00010165-M01, clone # 4F4, abnova) and anti-CAD (1:100, ab40800, abcam).
Rabinovich S., Adler L., Yizhak K., Sarver A., Silberman A., Agron S., Stettner N., Sun Q., Brandis A., Helbling D., Korman S., Itzkovitz S., Dimmock D., Ulitsky I., Nagamani S.C., Ruppin E, & Erez A. (2015). Diversion of aspartate in ASS1-deficient tumors fosters de novo pyrimidine synthesis. Nature, 527(7578), 379-383.
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