D0486

Manufactured by Agilent Technologies

Sourced in United Kingdom

The D0486 is a laboratory instrument designed for analytical applications. It provides precise measurements and data analysis capabilities. The detailed specifications and intended use of this product are not available at this time.

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Lab products found in correlation

Lsm 780 nlo, by Zeiss (1 mentions) Ab30917, by Abcam (1 mentions) Sta 011, by Cell Biolabs (1 mentions) P0217, by Agilent Technologies (1 mentions) Nupage novex 4 12 bis tris gel, by Thermo Fisher Scientific (1 mentions) Polyvinylidene difluoride transfer membrane, by GE Healthcare (1 mentions) Anti α tubulin, by Merck Group (1 mentions) Anti flag, by Cell Signaling Technology (1 mentions) Western lightning plus ecl, by PerkinElmer (1 mentions)

2 protocols using d0486

Quantification of Advanced Glycation End-Products

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Several measurements were performed on transverse slices of skin biopsies. Immunohistochemical staining was conducted with the anti-AGE antibodies anti-CML (ab30917, Abcam, Cambridge, UK) and anti-N^δ-(5-hydro-5-methyl-4-imidazolon-2-yl)-ornithine ([MG-H1], STA-011, cell biolabs, inc., NL) using goat anti-mouse IgG alkaline phosphatase (AP) conjugate ([Go-a-Mo-AP], D0486, Dako, Glostrup, DK) as chromogenic reporter. Histological localization of CML and MG-H1 was evaluated semi-quantitatively by two independent investigators (I.M.A and G.F.H.D). Hematoxylin and eosin (HE) stains were added to be able to identify individual cells.
Invasively assessed intrinsic fluorescence was measured by confocal microscopy using the ZEISS LSM 780 NLO (Zeiss, GER) and quantified by ImageJ. We used single (405nm and 440nm) and 2-photon (750nm, equivalent to 375nm) excitation, which corresponds to the excitation area of the AGE Reader.
In the second skin biopsy, including epidermis and dermis, the concentrations of CML, MG-H1 and pentosidine were assessed by ultra-performance liquid chromatography tandem mass spectrometry measurements (UPLC-MS/MS) and high-performance liquid chromatography (HPLC), respectively [20 , 21 (link), 22 (link)]. (Supplementary material).

Atzeni I.M., Boersema J., Pas H.H., Diercks G.F., Scheijen J.L., Schalkwijk C.G., Mulder D.J., van der Zee P, & Smit A.J. (2020). Is skin autofluorescence (SAF) representative of dermal advanced glycation endproducts (AGEs) in dark skin? A pilot study. Heliyon, 6(11), e05364.

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Western Blot Protein Detection Protocol

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Briefly, 20-50 µg of total protein extracts per sample were loaded on a precast NuPAGE® Novex 4-12% Bis-Tris Gel (Invitrogen) and subsequently blotted onto a polyvinylidene difluoride transfer membrane (GE Healthcare, Uppsala, Sweden). Membranes were blocked with 5% non-fatty dry milk (in 0.1% TBST on room temperature for 30 minutes followed by incubation with the primary antibody overnight at 4°C. A anti-CCND1 (1:10,000; Abcam, ab134175), anti-α-Tubulin (1:10000; Sigma, T9206) and anti-Flag (1:5000; Cell Signaling Technology, 14793S) antibody were used to visualize the proteins by immunodetection combined with chemiluminescence (Western Lightning Plus-ECL, PerkinElmer). As secondary antibody, a swine anti-rabbit (1:2500; Dako, P0217) and rabbit anti-mouse (1:10000; Dako, D0486) IgG antibody conjugated to horseradish peroxidase was used.

Van den Broeck T., Moris L., Gevaert T., Davicioni E., Boeckx B., Lambrechts D., Helsen C., Handle F., Ghesquière B., Soenen S., Smeets E., Eerlings R., El Kharraz S., Devlies W., Karnes R.J., Lotan T., Van Poppel H., Joniau S, & Claessens F. (2022). Antizyme Inhibitor 1 Regulates Matrikine Expression and Enhances the Metastatic Potential of Aggressive Primary Prostate Cancer. Molecular cancer research : MCR, 20(4).

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