Ab96032

Manufactured by Abcam

Sourced in United Kingdom

Ab96032 is a laboratory equipment product. It is a device designed for use in scientific research and analysis. The core function of this product is to perform a specific task or measurement related to laboratory procedures. No further details are provided to ensure an unbiased and factual approach.

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Lab products found in correlation

6 protocols using ab96032

Comprehensive Protein Expression Analysis

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Calnexin, VEGFA, CD31, HIF-1 alpha, (AB0037, AB0063, AB0092, AB0112, Sicgen, Portugal), β-actin, α-tubulin (A5316, T6199, Sigma, USA), PPARγ, AKT, p-AKT(Ser473), VEGFR2, PI3K (#2443, #9272, #4058, #2479, #4249, Cell Signaling, USA), ANG-2, F4/80, Perilipin-A, GLUT4, p-IR(Y1361), GLO-I, C/EBPalpha, PGC1alpha, ANGPTL4, AT1, HIF-2 alpha (Ab8452, Ab74383, Ab3526, Ab65267, Ab60946, Ab96032, Ab40764, Ab191838, Ab2920, Ab9391, Ab8365, Abcam, UK), CD11c, CD206 (bs-2058R, bs-2664R Bioss, USA) TIE-2, IRβ (sc-324, sc-57342, SantaCruz Biotechnology, USA), CEL (KH025, TransGenic Inc, Japan).

Rodrigues T., Matafome P., Sereno J., Almeida J., Castelhano J., Gamas L., Neves C., Gonçalves S., Carvalho C., Arslanagic A., Wilcken E., Fonseca R., Simões I., Conde S.V., Castelo-Branco M, & Seiça R. (2017). Methylglyoxal-induced glycation changes adipose tissue vascular architecture, flow and expansion, leading to insulin resistance. Scientific Reports, 7, 1698.

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Western Blotting for Glyoxalase I Quantification

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Protein extracts (20 μg) were separated under denaturing conditions on precast 4% to 20% mini gels, transferred to a nitrocellulose membrane, and blocked with 5% non-fat dry milk. Membranes were incubated overnight with rabbit-anti-Glo1 (1 : 1000; ab96032, Abcam, Cambridge, UK) or for one hour with mouse-anti-actin (1 : 5000; MP Biomedicals, Eschwege, Germany). Membranes were subsequently incubated with the appropriate horseradish peroxidase conjugated secondary antibody (1 : 5000; Jackson ImmunoResearch Laboratories, Europe; Dianova, Hamburg, Germany). Immunoreactive proteins were visualized on X-ray films using enhanced chemiluminescence detection reagents, according to the manufacturer's instructions. Densitometric analysis (ImageJ) was used to determine relative Glo1 levels, normalized to actin as a loading control.

Wortmann M., Hakimi M., Fleming T., Peters A.S., Sijmonsma T.P., Herzig S., Nawroth P.P., Böckler D, & Dihlmann S. (2015). A Glyoxalase-1 Knockdown Does Not Have Major Short Term Effects on Energy Expenditure and Atherosclerosis in Mice. Journal of Diabetes Research, 2016, 2981639.

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Immunohistochemical Analysis of Glyoxalase I

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Serial 2 μm transversal sections were prepared from formalin-fixed and paraffin-embedded aortic sinus specimens. After deparaffinization and rehydration, sections were pretreated in target retrieval solution pH 6.0 (DAKO, Glostrup, Denmark) in a steamer for 30 min. After 5 min washing in TBS buffer (20 mM Tris, 137 mM NaCl, pH 7.6) sections were incubated overnight with anti-Glo1 (1 : 300; ab96032, Abcam, Cambridge, UK). The DAKO real detection system peroxidase/AEC Rabbit/Mouse (DAKO, Glostrup, Denmark), containing biotinylated secondary antibody, streptavidin-HRP, and AEC/H2O2 substrate, was applied for further treatment, following the manufacturer's recommendation. Counterstaining was performed for 30 sec with Mayer's hemalum solution (Merck Darmstadt, Germany).

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Glutathione Metabolic Enzyme Assays

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Assay kits for determination of glutathione concentration (703302), and the activities of glutathione peroxidase (GPx) (703102), glutathione S-transferase (GST) (703302) and glutathione reductase (GR) (703202) were purchased from Cayman Chemical (Ann Arbor, MI). Antibodies against human GPx1 (ab22604), GST-ρi (ab134934), GR (ab55075), glutamate cysteine ligase catalytic subunit (GCLC) (ab55435), glyoxalase I (Glo1) (ab96032), advanced glycation end products (AGEs) (ab176173) and catalase (ab16731) were purchased from Abcam (Cambridge, MA), while the antibodies against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (sc305062), glucose-6-phosphate dehydrogenase (G6PD) (sc373887) and horseradish peroxidase-conjugated secondary antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Histopaque (10771) and assay kits for determination of glucose (GAGO-20), cholesterol (MAK043) and triglyceride (TR0100) concentrations were obtained from Sigma-Aldrich (St. Louis, MO).

Shan G., Yang F., Zhou L., Tang T., Okoro E.U., Yang H, & Guo Z. (2015). Increase in Blood Glutathione and Erythrocyte Proteins Related to Glutathione Generation, Reduction and Utilization in African-American Old Women with Diabetes. Journal of science, technology and environment, 5(1), 3000251.

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Antioxidant Enzyme and Glycation Marker Analysis

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Salts and organic solvents used in this study were all purchased from Sigma-Aldrich/Merck Portugal (Oeiras, Portugal) and Fischer Scientific (Pittsburgh, PA, USA). Antibodies were used to target catalase, GLO-1 (Ab76110, Ab96032, Abcam, Cambridge, UK), CML (KH024, TransGenic Inc, Tokyo, Japan), MG-H1 (HM5017, Hycult Biotech, Uden, Netherlands) and argpyrimidine (AGE06B, Nordic-MUbio, Susteren, the Netherlands). Calnexin (AB0037, Sicgen, Cantanhede, Portugal) was used as loading control.

Monteiro-Alfredo T., Caramelo B., Arbeláez D., Amaro A., Barra C., Silva D., Oliveira S., Seiça R, & Matafome P. (2021). Distinct Impact of Natural Sugars from Fruit Juices and Added Sugars on Caloric Intake, Body Weight, Glycaemia, Oxidative Stress and Glycation in Diabetic Rats. Nutrients, 13(9), 2956.

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Molecular Mechanisms of Metabolic Regulation

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Salts and organic solvents used in this study were all purchased from Lonza, Sigma-Aldrich/Merck, Alfa-Aesar, Fischer Scientifics and Panreac. Antibodies used were targeted to Catalase, Glo-1, GLUT2 (ab76110, ab96032, ab54460 Abcam, Cambridge, UK), GLUT4, PPARgamma, Insulin Receptor, AMPK, phospho-AMPK-Thr-172, Sirt1, phospho-Sirt1-Ser47 (#2213S, #2443S, #3025S, #2532S, #2535S, #9475S, #2314S, Cell Signaling Technology, Danvers, MA, USA) NRF2 (sc-518036, Santa Cruz Biotechnology, Dallas, TX, USA) phospho-NRF2 (Ser40) (PA5-67520, Invitrogen, Waltham, MA, USA). Calnexin and GAPDH (AB0037, AB0049-20, Sicgen, Carcavelos, Portugal) were used as loading control.

Monteiro-Alfredo T., Oliveira S., Amaro A., Rosendo-Silva D., Antunes K., Pires A.S., Teixo R., Abrantes A.M., Botelho M.F., Castelo-Branco M., Seiça R., Silva S., de Picoli Souza K, & Matafome P. (2021). Hypoglycaemic and Antioxidant Properties of Acrocomia aculeata (Jacq.) Lodd Ex Mart. Extract Are Associated with Better Vascular Function of Type 2 Diabetic Rats. Nutrients, 13(8), 2856.

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