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M0879

Manufactured by Agilent Technologies
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Sourced in Denmark, United States, Germany

The M0879 is a precision electronic instrument designed for analytical and scientific applications. It provides accurate measurements and data analysis capabilities required for various laboratory procedures. The core function of the M0879 is to perform high-quality data acquisition and processing tasks within the laboratory environment.

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Lab products found in correlation

Proliferating cell nuclear antigen (pcna), by Agilent Technologies (2 mentions) M0851, by Agilent Technologies (2 mentions) A5441, by Merck Group (2 mentions) Pab18350, by Abnova (2 mentions) K4063, by Agilent Technologies (2 mentions) Vectastain elite abc system, by Vector Laboratories (2 mentions) Ab16667, by Abcam (2 mentions) Ab8129, by Abcam (1 mentions) Hrp conjugated secondary antibody, by Nichirei Biosciences (1 mentions) Image pro plus 6, by Media Cybernetics (1 mentions) Dako envision dual link system hrp, by Agilent Technologies (1 mentions) Ab28364, by Abcam (1 mentions) Ab2349, by Abcam (1 mentions) Af4117, by R&D Systems (1 mentions) Ab5694, by Abcam (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Ab15580, by Agilent Technologies (1 mentions) 3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions) P smad 2 3 sc 11769, by Santa Cruz Biotechnology (1 mentions) Apoptag kit, by Merck Group (1 mentions)

25 protocols using m0879

1

Proliferating Cell Nuclear Antigen Immunohistochemistry

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Procedure was the same as Ki67 with antigen retrieval. The primary antibody used was proliferating cell nuclear antigen at a 1:100 concentration. (PCNA, DAKO; M0879) and the secondary antibody used was Alexa-Fluor 594 goat anti-mouse IgG at a 1:200 concentration.
Welker A.M., Jaros B.D., An M, & Beattie C.E. (2017). Changes in tumor cell heterogeneity after chemotherapy treatment in a xenograft model of glioblastoma. Neuroscience, 356, 35-43.
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2

Immunohistochemical Analysis of Tumor Cell Proliferation

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Mammary tumors were fixed in 10% formalin, embedded in paraffin and sectioned at 4 μm thickness. Sections were incubated overnight at 4°C with antibody to proliferating cell nuclear antigen (PCNA) (1:4000; M 0879, Dako, Denmark), followed by incubation with biotinylated secondary antibody and avidin/biotin peroxidase complex. The sections were then stained with 3′-diaminobenzamine substrate and counterstained with Modified Harris Haematoxylin. Tumor sections from three different animals per treatment group were stained. Representative images were taken randomly and nuclear staining was quantified using an Aperio ScanScope (Vista, CA) by counting at least 15,000 cells per slide.
Das Gupta S., Patel M., Wahler J., Bak M.J., Wall B., Lee M.J., Lin Y., Shih W.J., Cai L., Yang C.S, & Suh N. (2017). Differential gene regulation and tumor inhibitory activities of alpha-, delta- and gamma-tocopherols in estrogen-mediated mammary carcinogenesis. Cancer prevention research (Philadelphia, Pa.), 10(12), 694-703.
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3

Evaluating Breast Tissue Changes with Image-Guided Biopsies

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Image guided breast biopsies of mammographically dense tissue were performed at baseline and after 12 months. Five core specimens were obtained under local anesthesia at each time point. At the follow-up biopsy visit, the radiologist localized the previous biopsy site (biopsy clip placed at baseline) so that tissue was obtained from the same area of the breast. One core biopsy sample was formalin-fixed, paraffin-embedded, and examined for pathologic abnormalities. Immunohistochemistry (IHC) staining of breast tissue was performed for PCNA (Invitrogen) and TFF1 (Histostain Plus Kit from Zymed/Invitrogen) according to the manufacturer's instructions. Tissue was exposed to primary antibodies for PCNA (1:14000, Dako, M0879) and for TFF1(1:50). Negative controls for both stains were included. TFF1 was assessed by both intensity of stain (0 to 3+) and percent of cells with any staining. PCNA was assessed by the percent of positive epithelial cells in the ductal/lobular component of the breast tissue and the intensity of the strain (0 to +3). The pathologist (B.K.) was blinded to the time of biopsy.
Gatti-Mays M., Venzon D., Galbo C., Singer A., Reynolds J., Makariou E., Kallakury B., Heckman-Stoddard B., Korde L., Isaacs C., Warren R., Gallagher A, & Eng-Wong J. (2016). Exemestane use in postmenopausal women at high risk for invasive breast cancer: evaluating biomarkers of efficacy and safety. Cancer prevention research (Philadelphia, Pa.), 9(3), 225-233.
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4

Immunohistochemical Analysis of Mammary Tumors

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Mammary tumors were fixed in 10% formalin, embedded in paraffin and sectioned at 4 μm thickness. Sections were incubated overnight at 4°C with antibody to proliferating cell nuclear antigen (PCNA) (1:8000; M 0879, Dako, Denmark), followed by incubation with biotinylated secondary antibody and avidin/biotin peroxidase complex. The sections were then stained with 3′-diaminobenzamine substrate and counterstained with Modified Harris Haematoxylin. Tumor sections from three different animals per treatment group were stained. Representative images (40X magnification) were taken randomly and staining density was quantified using an Aperio ScanScope (Vista, CA) by counting at least 15,000 cells per slide.
Das Gupta S., Sae-tan S., Wahler J., So J.Y., Bak M.J., Cheng L.C., Lee M.J., Lin Y., Shih W.J., Shull J.D., Safe S., Yang C.S, & Suh N. (2015). Dietary γ-Tocopherol Rich Mixture Inhibits Estrogen-Induced Mammary Tumorigenesis by Modulating Estrogen Metabolism, Antioxidant Response and PPARγ. Cancer prevention research (Philadelphia, Pa.), 8(9), 807-816.
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5

Immunohistochemistry of Subcutaneous Tumors

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Paraffin-embedded subcutaneous tumor tissues were sectioned (5 μM thick) and immunostained with pSTAT3, proliferating cell nuclear antigen (PCNA), Iba-1, CD3, CD34, and CD204. Anti-pSTAT3 (D3A7, Cell Signaling Technology; 1:200), anti-PCNA (M0879, DAKO; 1:200), anti-Iba-1 (019-19741, Wako; 1:4000), anti-CD3 (413591, Nichirei Biosciences; 1:2), anti-CD34 (ab8129, Abcam; 1:2000), and anti-CD204 (2F8, Invitrogen; 1:2000) antibodies were used as primary antibodies. The sections were subsequently treated with an HRP-conjugated secondary antibody (Nichirei, Tokyo, Japan), followed by the visualization with diaminobenzidine.
Pan C., Fujiwara Y., Horlad H., Shiraishi D., Iriki T., Tsuboki J., Ikeda T, & Komohara Y. (2020). Flavonoid Compounds Contained in Epimedii Herba Inhibit Tumor Progression by Suppressing STAT3 Activation in the Tumor Microenvironment. Frontiers in Pharmacology, 11, 262.
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6

Immunohistochemical Analysis of Vascular Markers

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Tissue sections were de-paraffined using xylene and a graded series of alcohols, non-specific background staining of endogenous peroxidase was treated with 1% hydrogen peroxide for 30 minutes, and sections were blocked with 1% bovine serum albumin prior to incubation with primary antibodies overnight at 4°C. Primary antibodies were directed against the endothelial marker CD31 (ab28364, Abcam; 1:50 dilution), the smooth muscle marker α-actin (ab5694, Abcam; 1:100 dilution), the progenitor marker CD34 (AF4117, R&D; 1:100 dilution), the endothelial marker VEGFR2 (ab2349, Abcam; 1:50 dilution), the M2 macrophage marker anti-transglutaminase 2 (TGM2; #37557; Cell Signaling; 1:50 dilution), proliferating cell nuclear antigen (PCNA; M0879, Dako ; 1:100 dilution) or the apoptosis marker cleaved caspase-3 (#9661; Cell Signaling; 1:100 dilution). Detection was performed using Dako EnVision™ + Dual Link System-HRP (Dako; Carpinteria, CA), and counterstained with Mayer’s Hematoxylin. The integrated optical density of smooth muscle cells (SMC) in the neointima were analyzed using Image-Pro Plus 6.0 software (Media Cybernetics; Rockville, MD). Cells staining positively for CD34, VEGFR2, PCNA (proliferation index), or cleaved caspase-3 (apoptosis index) were directly counted in 4 high power fields, and compared to the total number of cells in the field.
Bai H., Kuwahara G., Wang M., Brownson K., Foster T., Yamamoto K., Xing Y, & Dardik A. (2014). Pretreatment of pericardial patches with antibiotics does not alter patch healing in vivo. Journal of vascular surgery, 63(4), 1063-1073.
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7

Immunohistochemical Analysis of Proliferation Markers

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The expression of proliferation- associated antigens Ki67 and proliferating cell nuclear antigen (PCNA) in tissues was measured by immunohistochemistry assay. After conventional paraffin embedding, sectioning, dewaxing and hydration, the tumor tissues were treated with antigen repair buffer (pH 6.0). Following treatment with 0.3% H2O2 for 20 min, the tumor tissue sections were blocked with 5% BSA (Sigma-Aldrich; Merck KGaA) and incubated with primary antibodies (anti-Ki-67, Abcam, ab15580, 1:200 dilution; anti-PCNA, Dako, M0879, 1:50 dilution) overnight at 4°C. On the following day, the tissue sections were washed with PBS three times and incubated with secondary antibodies at 37°C for 1 h, followed by treatment with DAB chromogen.
Zhang R., Zhu H., Yuan Y., Wang Y, & Tian Z. (2020). SPAG6 promotes cell proliferation and inhibits apoptosis through the PTEN/PI3K/AKT pathway in Burkitt lymphoma. Oncology Reports, 44(5), 2021-2030.
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8

Immunohistochemical Staining for Kidney Analysis

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For immunohistochemical staining kidneys were fixed in methyl carnoy’s solution and embedded in paraffin. 2μm sections were stained as described below.
For PCNA and survivin immunostaining kidney tissue was pretreated with TRS (DAKO, Hamburg, Germany). Sections were blocked with 3% H2O2. Primary antibodies were incubated overnight at the following dilutions: PCNA for proliferating cells (M0879; DAKO) 1:500, survivin (AF886; R&D Systems) 1:250, F4/80 for macrophages (LMU8949; Linaris, Dossenheim, Germany) 1:50, CD3 for T-cells (I7A2; BioLegend) 1:300, CD4 for T-helper cells (14-9766-82; eBioscience) 1:200, CD8a for cytotoxic T-cells (14-0808-82; eBioscience) 1:200, vimentin (GP53; Progen, Heidelberg, Germany) 1:50, p-SMAD2/3 (sc-11769; Santa Cruz Biotechnologies) 1:5000, α-smooth muscle actin (M 0851; DAKO) 1:50. Appropriate secondary antibodies (Vector, Burlingame, CA) were diluted 1:500, before avidin D peroxidase (Vector) was applied at a dilution of 1:2000. Finally DAB (Vector) was added, sections were counterstained with hematoxylin and covered with entellan.
Marek I., Lichtneger T., Cordasic N., Hilgers K.F., Volkert G., Fahlbusch F., Rascher W., Hartner A, & Menendez-Castro C. (2016). Alpha8 Integrin (Itga8) Signalling Attenuates Chronic Renal Interstitial Fibrosis by Reducing Fibroblast Activation, Not by Interfering with Regulation of Cell Turnover. PLoS ONE, 11(3), e0150471.
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9

Evaluating Tumor Response to Treatments

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PBS, LDH, MTX, or MTX-LDH was systemically administered to mice bearing MCF-7/mot orthotopic tumors. Six days after the third treatment, representative tumor tissue was collected from each group, fixed in 10% formalin, embedded in paraffin (Wax-it, Vancouver, Canada), and cut into 5-μm thick sections. Representative sections were stained with hematoxylin, eosin, and with an antibody specific for proliferating cellular nuclear antigen (M0879, Dakocytomation, Carpinteria, CA). Apoptotic cells were identified by the terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) assay using an in situ ApopTag kit (Chemicon International, Temecula, CA).
Choi G., Kwon O.J., Oh Y., Yun C.O, & Choy J.H. (2014). Inorganic Nanovehicle Targets Tumor in an Orthotopic Breast Cancer Model. Scientific Reports, 4, 4430.
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10

Immunohistochemical Analysis of Mortalin, p53, and PCNA

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Formaldehyde-fixed tissues were transferred to a paraffin-embedded block, sectioned at 4-μm thicknesses. After tissue deparaffinization and rehydration, endogenous peroxidase activity was blocked by 10-min incubation at room temperature with absolute methanol containing 1% hydrogen peroxide. The tissue sections were incubated with a primary antibody against mouse anti-mortalin monoclonal antibody (C1-3), rabbit anti-p53 (sc-6243; Santa Cruz biotechnology, Santa Cruz, CA), and mouse anti-PCNA (M0879; DAKO, Carpinteria, CA) at 4 °C overnight. After incubation with the secondary antibody (Super Sensitive™ Polymer-HRP IHC, BioGenex) for 1 h at room temperature, the bound complexes were visualized by incubating tissue sections with 0.05% diaminobenzidine and 0.003% hydrogen peroxide. The sections were counterstained with Harris hematoxylin and then dehydrated and mounted. Mortalin, p53, and PCNA protein levels were semi-quantitatively analyzed using MetaMorph® image analysis software (Universal Image Corp., Buckinghamshire, UK). Results are expressed as mean optical density of six different digital images per sample.
Lee W.J., Ahn H.M., Na Y., Wadhwa R., Hong J, & Yun C.O. (2017). Mortalin deficiency suppresses fibrosis and induces apoptosis in keloid spheroids. Scientific Reports, 7, 12957.
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