Ab74383

Manufactured by Abcam

Sourced in United Kingdom

Ab74383 is a laboratory product manufactured by Abcam. It is a piece of lab equipment used for scientific research purposes. No further details about its core function or intended use are provided.

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Lab products found in correlation

7 protocols using ab74383

Antibody-Based Analysis of Metabolic Signaling

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Salts and organic solvents used in solution preparations were purchased from Fisher Scientific (Leicestershire, UK), Sigma Chemicals (USA), or Merck Darmstadt (Germany), with the highest grade of purity commercially available. Antibodies used were targeted to AMPK and (Thr172)AMPK (#2532, #2535, Cell Signaling, USA); F4/80, GLUT2, ACC, (Ser79)ACC, and (Tyr1163)IRβ (ab74383, ab54460, ab72046, ab68191, and Ab60946, Abcam, UK); IRβ (sc-57342, Santa Cruz Biotechnology, USA) argpyrimidine (AGE06B, Nordic MUbio, Netherlands); and MG-H1 (HM5017, Hycult Biotech, Netherlands). Calnexin was used as loading control (AB0037, SICGEN, Portugal).

Neves C., Rodrigues T., Sereno J., Simões C., Castelhano J., Gonçalves J., Bento G., Gonçalves S., Seiça R., Domingues M.R., Castelo-Branco M, & Matafome P. (2019). Dietary Glycotoxins Impair Hepatic Lipidemic Profile in Diet-Induced Obese Rats Causing Hepatic Oxidative Stress and Insulin Resistance. Oxidative Medicine and Cellular Longevity, 2019, 6362910.

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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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Macrophage Identification via F4/80

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Immunohistochemistry was performed with a rabbit primary antibody for macrophage F4/80 (ab74383, Abcam). Ventricular sections were heat retrieved with a 10mM Na-citrate buffer (pH 8.5) and incubated with the primary antibody overnight at 4°C. Anti-rabbit secondary antibodies were used for detection with 3,3′-diaminobenzidine chromagen (Vector Labs).

Younce C.W., Niu J., Ayala J., Burmeister M.A., Smith L.H., Kolattukudy P, & Ayala J.E. (2014). Exendin-4 Improves Cardiac Function in Mice Overexpressing Monocyte Chemoattractant Protein-1 in Cardiomyocytes. Journal of molecular and cellular cardiology, 0, 172-176.

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Immunohistochemical Analysis of Liver Tissue

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Liver tissues from median and left lobes were collected, incubated in 4% paraformaldehyde (MPX00553, Thermofisher Scientific, Rockford, IL) and embedded in paraffin wax according to the traditional method. Sections were made at 4 μm thickness and stained with hematoxylin-eosin (H&E) for observation under a light microscope.
To perform immunohistochemistry staining, liver sections were prepared by heating at 70°C for 30 min. Having dewaxed in dimethylbenzene for 10 min, we dehydrated the sections in a graded series of alcohol. Antigen retrieval was then performed by microwaving the samples in citrate buffer for four cycles. Endogenous peroxidase activity was blocked by treatment with 3% H₂O₂ for 15 min at room temperature. Nonspecific proteins were blocked with 5% bovine serum albumin (BSA) (BP9706100, Thermofisher Scientific) for 30 min. The sections were incubated with an F4/80 (ab74383, 1:200, Abcam Company, Cambridge, MA) antibody at 4°C overnight. The sections were then washed with phosphate-buffered saline (PBS) and treated immediately with a corresponding secondary antibody at 1:500 in PBS (sc2027, Santa Cruz Biotechnology) for another 30 min. The positive reactions were finally visualized using a diaminobenzidine (DAB) kit (ab64238, Abcam) and the stained tissue was then observed under a light microscope.

Zhuang Y., Li Y., Li X., Xie Q, & Wu M. (2016). Atg7 Knockdown Augments Concanavalin A-Induced Acute Hepatitis through an ROS-Mediated p38/MAPK Pathway. PLoS ONE, 11(3), e0149754.

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Immunofluorescence Staining for Macrophage and Proliferation Analysis

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The immunofluorescence staining experiments were performed, and the staining results were analyzed by two investigators (L.G. and Y.Z.) who were blinded to the results of flow cytometry, in vivo imaging, and tumor therapy. For CD206 and F4/80 analyses, frozen sections were incubated with rat anti-mouse CD206 (1:100; clone C068C2, IgG2a; Biolegend, San Diego, CA) and rabbit anti-mouse F4/80 antibodies (1:100; ab74383; Abcam, Cambridge, MA) and then visualized using Dylight549- or FITC-conjugated secondary antibodies (1:200; Earthox, Millbrae, CA) under a confocal microscope (Leica, Wetzlar, Germany). For Ki67 analysis, frozen sections were incubated with rabbit anti-mouse Ki67 (1:100; AB9260; Millipore, Billerica, MA) and then visualized using a Dylight549-conjugated secondary antibody (1:200; Earthox, Millbrae, CA). The tumor cell proliferation index was calculated as the percentage of Ki67-positive nuclei among the total number of nuclei.

Zhang C., Yu X., Gao L., Zhao Y., Lai J., Lu D., Bao R., Jia B., Zhong L., Wang F, & Liu Z. (2017). Noninvasive Imaging of CD206-Positive M2 Macrophages as an Early Biomarker for Post-Chemotherapy Tumor Relapse and Lymph Node Metastasis. Theranostics, 7(17), 4276-4288.

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Macrophage Identification in Renal Tissue

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Immunohistochemistry was performed with an anti-F4/80 antibody (ab74383; Abcam) to identify infiltrated macrophages in renal tissue.

Wang F., Zhang G., Lu Z., Geurts A.M., Usa K., Jacob H.J., Cowley A.W., Wang N, & Liang M. (2015). Antithrombin III/SerpinC1 insufficiency exacerbates renal ischemia/reperfusion injury. Kidney International, 88(4), 796-803.

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Macrophage Infiltration in Renal Tissue

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Immunohistochemistry was performed with an anti-F4/80 antibody (Abcam, #ab74383) to identify infiltrated macrophages in renal tissue.

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