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Ab28455

Manufactured by Abcam
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Sourced in China

Ab28455 is a lab equipment product manufactured by Abcam. It is a device used for analytical purposes in research and laboratory settings. The core function of this product is to perform specific tasks or measurements required for scientific investigations, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Ab83690, by Abcam (2 mentions) Ab37583, by Abcam (2 mentions) Nb100 122, by Novus Biologicals (2 mentions) Nb100 134, by Novus Biologicals (2 mentions) Rabbit 2729, by Cell Signaling Technology (2 mentions) Mouse 5415, by Cell Signaling Technology (2 mentions) Sab1405798, by Merck Group (2 mentions) Hpa005857, by Merck Group (2 mentions) Dichlorofluorescin diacetate, by Beyotime (2 mentions) Ab70359, by Abcam (2 mentions) Ab52771, by Abcam (2 mentions) Phospho ampkα thr172, by Cell Signaling Technology (2 mentions) Sodium pyruvate, by Thermo Fisher Scientific (2 mentions) Phospho c jun ser73, by Cell Signaling Technology (2 mentions) N acetylcysteine nac, by Merck Group (2 mentions) Phospho acc ser79, by Cell Signaling Technology (2 mentions) Z vad fmk, by Santa Cruz Biotechnology (2 mentions) 3 mercaptopicolinic acid, by Santa Cruz Biotechnology (2 mentions) Dimethyl 2 oxoglutarate, by Tokyo Chemical Industry (2 mentions) Dimethyl fumarate, by Tokyo Chemical Industry (2 mentions)

7 protocols using ab28455

1

Antibody Detection of Metabolic Proteins

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Antibodies detecting HDAC1 (5356), HSP90 (4877), GST (2625), HA (3724), and normal IgG isotopic controls (rabbit-2729, mouse-5415) were purchased from Cell Signaling Technology. Antibodies detecting HIF1α (NB100-134) and HIF2α (NB100-122) were obtained from Novus, and antibodies against PCK1 (ab28455), G6PC (ab83690), and PFKL (ab37583) were from Abcam. D-fructose-1, 6-bisphosphate trisodium salt (47810), 2-hydroxyethyl agarose (A4018), and antibodies detecting FBP1 (SAB1405798 and HPA005857) were purchased from Sigma.
Li B., Qiu B., Lee D.S., Walton Z.E., Ochocki J.D., Mathew L.K., Mancuso A., Gade T.P., Keith B., Nissim I, & Simon M.C. (2014). Fructose-1, 6-bisphosphatase opposes renal carcinoma progression. Nature, 513(7517), 251-255.
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2

Western Blot Analysis of Liver Proteins

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Total proteins were extracted from the frozen liver tissues using the T-PER tissue protein extraction reagent (Thermo Fisher Scientific Inc., Waltham, MA), and used for western blotting. Immunoblotting was performed using the following primary antibodies: PCK1 (1:1000; ab28455, Abcam, Toronto, Canada), G6PC (ab83690; Abcam), FOXO1 (1:000; 2880, Cell Signaling Technology, Danvers, MA), and β-actin (A5216, Sigma‐Aldrich Co., St. Louis, MO), and incubated with anti- rabbit HRP-conjugated secondary antibody (1:10,000; NA934, GE Healthcare, Chicago, IL, USA) anti- mouse HRP (1:10,000; NA931, GE Healthcare). Immunoreactive bands were visualized using ECL (Thermo Fisher Scientific).
Kashiwagi Y., Aburaya S., Sugiyama N., Narukawa Y., Sakamoto Y., Takahashi M., Uemura H., Yamashita R., Tominaga S., Hayashi S., Nozaki T., Yamada S., Izumi Y., Kashiwagi A., Bamba T., Ishihama Y, & Murakami S. (2021). Porphyromonas gingivalis induces entero-hepatic metabolic derangements with alteration of gut microbiota in a type 2 diabetes mouse model. Scientific Reports, 11, 18398.
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3

Metabolic Regulation Pathway Investigation

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3-mercaptopicolinic acid, Z-VAD-FMK and Necrostatin-1 were purchased from Santa Cruz Biotechnology. Dimethyl-2-oxoglutarate, dimethyl-fumarate, dimethyl-succinate and dimethyl-malate were purchased from Tokyo Chemical Industry, Sodium pyruvate and L-glutamine were purchased from Thermo Fisher Scientific, N-acetylcysteine (NAC) was purchased from Sigma-Aldrich, and dichlorofluorescin diacetate from Beyotime.
Antibodies against PCK1 (ab28455, Abcam), PCK2 (ab70359, Abcam) and YAP1 (ab52771, Abcam), phospho-AMPKα (Thr172) (Cell Signaling Technology, 2535), AMPKα (Cell Signaling, 2532), phospho-ACC (Ser79) (Cell Signaling, 11818) and phospho-c-Jun (Ser73) (Cell Signaling, 3270) were purchased commercially.
pBABE-Flag-PCK1 and pQCXIH-Flag-PCK2 were cloned from cDNA provided by Dr. Jiahuai Han (Xiamen University, Xiamen, China). The PB[CMV-myc-YAP-5SA]DS, PB[Act-RFP]DS and Act-PB Transposase plasmids were generous gifts from Dr. Bin Zhao (Zhejiang University, Hangzhou, China). The PB[CMV-flag-PCK1]DS donor plasmids were constructed by excising Act-RFP from the PB[Act-RFP]DS plasmid and ligating the corresponding fragments excised from pQCXIH vectors.
Liu M.X., Jin L., Sun S.J., Liu P., Feng X., Cheng Z.L., Liu W.R., Guan K.L., Shi Y.H., Yuan H.X, & Xiong Y. (2018). Metabolic reprogramming by PCK1 promotes TCA cataplerosis, oxidative stress and apoptosis in liver cancer cells and suppresses hepatocellular carcinoma. Oncogene, 37(12), 1637-1653.
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4

Immunohistochemical Staining Protocol

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Hematoxylin–eosin (HE) and immunohistochemical staining were performed by standard techniques as previously described. The following primary antibodies were used: anti-CD34 (ab81289; Abcam), anti-NSE (PA5-27,452; Invitrogen), anti-D2-40 (MA1-83,884; Invitrogen), anti-CA9 (ab184006; Abcam), anti-PCK (ab28455; Abcam), anti-Oligo2 (ab109186; Abcam), anti-SOX9 (#82,630; Cell signaling technology), anti-Inhibin (PA5-81,202; Invitrogen), anti-GFAP (ab7260; Abcam), anti-S100 (ab183979; Abcam), and anti-Ki67 (ab15580; Abcam).
Wang Q., Liu W., Zhang S., Liang Z., Jiang L., Xue A., Cen X, & Bu Q. (2021). Combined transcriptomic and lipidomic analysis reveals aberrant lipid metabolism in central nervous system hemangioblastomas. Scientific Reports, 11, 1314.
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5

Antibody Detection of Metabolic Proteins

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Antibodies detecting HDAC1 (5356), HSP90 (4877), GST (2625), HA (3724), and normal IgG isotopic controls (rabbit-2729, mouse-5415) were purchased from Cell Signaling Technology. Antibodies detecting HIF1α (NB100-134) and HIF2α (NB100-122) were obtained from Novus, and antibodies against PCK1 (ab28455), G6PC (ab83690), and PFKL (ab37583) were from Abcam. D-fructose-1, 6-bisphosphate trisodium salt (47810), 2-hydroxyethyl agarose (A4018), and antibodies detecting FBP1 (SAB1405798 and HPA005857) were purchased from Sigma.
Li B., Qiu B., Lee D.S., Walton Z.E., Ochocki J.D., Mathew L.K., Mancuso A., Gade T.P., Keith B., Nissim I, & Simon M.C. (2014). Fructose-1, 6-bisphosphatase opposes renal carcinoma progression. Nature, 513(7517), 251-255.
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6

Validation of Dysregulated Proteins in ccRCC

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Two most significantly dysregulated proteins (PCK1 and SNRPF) were chosen to be confirmed by western blotting. Briefly, 20 µg of total protein were separated on a 10% SDS-PAGE gel. Proteins were then transferred to a PVDF membrane and probed with the following polyclonal antibodies: anti-PCK1 and anti-SNRPF (ab28455 and ab156587, 1:500; Abcam). GAPDH (sc-48166, 1:1,000; Santa Cruz Biotechnology, Inc.) was used as a loading control. Protein expression was visualized after incubation with secondary anti-rabbit antibodies conjugated with horseradish peroxidase and enhanced chemiluminescence reagent.
The intensity of protein staining was determined using Gel-Pro Analyzer 4.0. Log2 fold change (FC) in expression of the two proteins between four ccRCC tumor tissues and matched normal kidney tissues is presented as a graph.
Sun X., Zhang H., Luo L., Zhong K., Ma Y., Fan L., Fu D, & Wan L. (2016). Comparative proteomic profiling identifies potential prognostic factors for human clear cell renal cell carcinoma. Oncology Reports, 36(6), 3131-3138.
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7

Metabolic Regulation Pathway Investigation

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3-mercaptopicolinic acid, Z-VAD-FMK and Necrostatin-1 were purchased from Santa Cruz Biotechnology. Dimethyl-2-oxoglutarate, dimethyl-fumarate, dimethyl-succinate and dimethyl-malate were purchased from Tokyo Chemical Industry, Sodium pyruvate and L-glutamine were purchased from Thermo Fisher Scientific, N-acetylcysteine (NAC) was purchased from Sigma-Aldrich, and dichlorofluorescin diacetate from Beyotime.
Antibodies against PCK1 (ab28455, Abcam), PCK2 (ab70359, Abcam) and YAP1 (ab52771, Abcam), phospho-AMPKα (Thr172) (Cell Signaling Technology, 2535), AMPKα (Cell Signaling, 2532), phospho-ACC (Ser79) (Cell Signaling, 11818) and phospho-c-Jun (Ser73) (Cell Signaling, 3270) were purchased commercially.
pBABE-Flag-PCK1 and pQCXIH-Flag-PCK2 were cloned from cDNA provided by Dr. Jiahuai Han (Xiamen University, Xiamen, China). The PB[CMV-myc-YAP-5SA]DS, PB[Act-RFP]DS and Act-PB Transposase plasmids were generous gifts from Dr. Bin Zhao (Zhejiang University, Hangzhou, China). The PB[CMV-flag-PCK1]DS donor plasmids were constructed by excising Act-RFP from the PB[Act-RFP]DS plasmid and ligating the corresponding fragments excised from pQCXIH vectors.
Liu M.X., Jin L., Sun S.J., Liu P., Feng X., Cheng Z.L., Liu W.R., Guan K.L., Shi Y.H., Yuan H.X, & Xiong Y. (2018). Metabolic reprogramming by PCK1 promotes TCA cataplerosis, oxidative stress and apoptosis in liver cancer cells and suppresses hepatocellular carcinoma. Oncogene, 37(12), 1637-1653.
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