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Rmr622

Manufactured by Biocare Medical
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Sourced in Canada

The RMR622 is a laboratory centrifuge designed for general-purpose use. It has a maximum speed of 6,000 RPM and a rotor capacity of 6 x 50 mL tubes. The centrifuge is equipped with an imbalance detection system and an automatic rotor identification feature.

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Lab products found in correlation

Rodent block m, by Biocare Medical (3 mentions) Dab substrate, by Thermo Fisher Scientific (2 mentions) Mounting medium, by Thermo Fisher Scientific (2 mentions) Vim antibody, by Cell Signaling Technology (2 mentions) Mca1477t, by Bio-Rad (1 mentions) Ab6640, by Abcam (1 mentions) Ab16669, by Abcam (1 mentions) Ab81321, by Abcam (1 mentions) Ab179475, by Abcam (1 mentions) Mm620l, by Biocare Medical (1 mentions) Sf994, by Thermo Fisher Scientific (1 mentions) Xylene substitute, by Thermo Fisher Scientific (1 mentions) Decloaking chamber, by Biocare Medical (1 mentions) Methyl green, by Vector Laboratories (1 mentions) Da vinci green diluent, by Biocare Medical (1 mentions) Diva decloaker, by Biocare Medical (1 mentions) Cdh13, by Merck Group (1 mentions)

8 protocols using rmr622

1

Quantitative Analysis of T-Cell Infiltration

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Tumors were fixed in 10% formalin for 24 hours, washed in 70% ethanol for 24 hours, and embedded for histological analysis. Five-micron (5 μm) sections were cut from paraffin blocks. Briefly, all slides underwent deparaffinization and sequential antigen retrieval. Primary antibody against mouse CD3 (cat. #MCA1477T, Bio-Rad) and horse-radish peroxidase–coupled secondaries (cat. #RMR622, Biocare) were used according to the manufacturer's instructions. After whole-slide scanning at ×20 using Hamamatsu NanoZoomer whole-slide scanner, ImageJ was used for the quantifications of all IHC staining.
Hu H., Khodadadi-Jamayran A., Dolgalev I., Cho H., Badri S., Chiriboga L.A., Zeck B., Lopez De Rodas Gregorio M., Dowling C.M., Labbe K., Deng J., Chen T., Zhang H., Zappile P., Chen Z., Ueberheide B., Karatza A., Han H., Ranieri M., Tang S., Jour G., Osman I., Sucker A., Schadendorf D., Tsirigos A., Schalper K.A., Velcheti V., Huang H.Y., Jin Y., Ji H., Poirier J.T., Li F, & Wong K.K. (2021). Targeting the Atf7ip–Setdb1 Complex Augments Antitumor Immunity by Boosting Tumor Immunogenicity. Cancer Immunology Research, 9(11), 1298-1315.
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2

Immunohistochemical Analysis of Tumor Samples

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Tumors were fixed in formalin for 24 hours, paraffin embedded, sectioned and stained according to standard procedures. Briefly, slides were baked at 60°C for 1 hour then deparaffinized and rehydrated. After antigen retrieval in citrate buffer, endogenous peroxidases were inactivated by 3% hydrogen peroxide. Non-specific signals were blocked using Rodent Block M (Biocare RBM961). Then slides were stained using VIM antibody (Cell Signaling; 5741) overnight at 4°C. After overnight incubation, the slides were washed and incubated with secondary antibody (HRP-polymers, Biocare RMR622) for 30 min at room temperature. The slides were washed and stained with DAB substrate (ThermoFisher Scientific). The slides were then counterstained with hematoxylin and mounted with mounting medium (Richard-Allan Scientific). Cells were counted using imageJ based on a minimum of 9 20x images from each sample. P values were calculated using a wilcoxon test.
Terranova C.J., Tang M., Maitituoheti M., Raman A.T., Ghosh A.K., Schulz J., Amin S.B., Orouji E., Tomczak K., Sarkar S., Oba J., Creasy C., Wu C.J., Khan S., Lazcano R., Wani K., Singh A., Barrodia P., Zhao D., Chen K., Haydu L.E., Wang W.L., Lazar A.J., Woodman S.E., Bernatchez C, & Rai K. (2021). Reprogramming of bivalent chromatin states in NRAS mutant melanoma suggests PRC2 inhibition as a therapeutic strategy. Cell reports, 36(3), 109410.
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3

Immune Cell Quantification in Myocardium

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Immunostaining was performed on mid-ventricular sections to measure T-lymphocyte and macrophage infiltration, respectively using antibody against CD3 (#ab 16669) and F4/80 (#ab 6640) (Abcam, Cambridge, United Kingdom). Detection kits RMR622 (for CD3) and RT517 (for F4/80) (BioCare Medical, Pacheco, CA) were used for detecting positive cells, in which HRP-conjugated secondary antibodies were reacted with DAB as the chromogen. Nuclei were counterstained with hematoxylin.
Mohammadzadeh N., Lunde I.G., Andenæs K., Strand M.E., Aronsen J.M., Skrbic B., Marstein H.S., Bandlien C., Nygård S., Gorham J., Sjaastad I., Chakravarti S., Christensen G., Engebretsen K.V, & Tønnessen T. (2019). The extracellular matrix proteoglycan lumican improves survival and counteracts cardiac dilatation and failure in mice subjected to pressure overload. Scientific Reports, 9, 9206.
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4

Immunohistochemistry and TUNEL Assay for TRIP13 in Mouse Kidneys

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Paraffin embedded wild-type Trip13+/+ mouse kidneys were sectioned (4 μm), and epitope retrieval was performed using a steamer bath. TRIP13 antibody (1:800 dilution) was applied overnight at 4 °C. Secondary rabbit‐on‐rodent HRP polymer (Biocare Medical Catalog #RMR622; Concord, CA) was incubated for 1 hour at room temperature. DAB was added in the presence of hydrogen peroxide until brown color was developed.
Immunostaining with injured and uninjured (contralateral) kidneys from the WT Trip13+/+ and mutant Trip13Gt/Gtmice were performed for TUNEL (apoptosis marker) using similar protocols as previously described in our lab66 (link). All sections were scanned using Aperio ImageScope software and images were counted for TUNEL‐positive cells out of a total of 1,000 nuclei from images taken at 40X magnification.
Pressly J.D., Hama T., Brien S.O., Regner K.R, & Park F. (2017). TRIP13-deficient tubular epithelial cells are susceptible to apoptosis following acute kidney injury. Scientific Reports, 7, 43196.
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5

Quantitative Immunohistochemical Analysis of PDAC

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PDAC tumor microarrays (US Biomax, slide PA242c) were stained with anti-NRP-1 (Abcam ab81321) or anti-alpha v integrin (Abcam ab179475) primaries in accordance with manufacturer instructions, followed by HRP secondaries (BioCare Rabbit-on-Rodent RMR622 and Mouse-on-Mouse MM620L polymers). Slides were digitized using an Aperio slide scanner and quantified using standard DAB and hematoxylin deconvolution functions in ImageJ. Grading was objective and based on linearly spaced bins by DAB to hematoxylin ratio (Grade 1: 0-5; Grade 2: 5-10, Grade 3: 10-15, Grade 4: 15+).
Lo J.H., Hao L., Muzumdar M.D., Raghavan S., Kwon E.J., Pulver E.M., Hsu F., Aguirre A.J., Wolpin B.M., Fuchs C.S., Hahn W.C., Jacks T, & Bhatia S.N. (2018). iRGD-guided tumor penetrating nanocomplexes for therapeutic siRNA delivery to pancreatic cancer. Molecular cancer therapeutics, 17(11), 2377-2388.
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6

Immunohistochemical Analysis of CD34+ Cells

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Tissues were dissected and fixed in 10% neutral buffered formalin (Fisher Scientific, SF99-4) for 24 hours at room temperature and processed in the Hypercenter XP (Thermo Fisher) using a standard dehydration protocol before paraffin embedding and microtome sectioning into 4 μm sections. Tissue sections were deparaffinized with xylene substitute (Thermo Fisher) before rehydration with gradients of alcohol. Heat-induced antigen retrieval was performed in Rodent Decloaker solution (Biocare Medical, RD913) in the Decloaking chamber (Biocare Medical). Non-specific interaction of primary antibodies with the tissue was blocked by Rodent block M (Biocare Medical, RBM961). Tissues were then stained with Rabbit anti-CD34 (Abcam, ab81289) at 1:3000 dilution in Da Vinci Green diluent (Biocare Medical, PD900) overnight at 4° C. The tissue section was stained with Rabbit-on-rodent-HRP polymer (Biocare Medical, RMR622). The slides were treated with Vector ImmPact® VIP peroxidase substrate (Vector labs, SK-4605) for 10 minutes at room temperature. Methyl green (Vector labs, H-3402) was used to counterstain.
Shibad V., Bootwala A., Mao C., Bader H., Vo H., Landesman-Bollag E., Guo C., Rubio A., Near R., Gao W., Challa S., Chukka V., Gao J., Kelly A., Landesman T., VanHelene T, & Zhong X. (2021). L2pB1 Cells Contribute to Tumor Growth Inhibition. Frontiers in Immunology, 12, 722451.
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7

Immunohistochemical Analysis of Tumor Samples

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Tumors were fixed in formalin for 24 hours, paraffin embedded, sectioned and stained according to standard procedures. Briefly, slides were baked at 60°C for 1 hour then deparaffinized and rehydrated. After antigen retrieval in citrate buffer, endogenous peroxidases were inactivated by 3% hydrogen peroxide. Non-specific signals were blocked using Rodent Block M (Biocare RBM961). Then slides were stained using VIM antibody (Cell Signaling; 5741) overnight at 4°C. After overnight incubation, the slides were washed and incubated with secondary antibody (HRP-polymers, Biocare RMR622) for 30 min at room temperature. The slides were washed and stained with DAB substrate (ThermoFisher Scientific). The slides were then counterstained with hematoxylin and mounted with mounting medium (Richard-Allan Scientific). Cells were counted using imageJ based on a minimum of 9 20x images from each sample. P values were calculated using a wilcoxon test.
Terranova C.J., Tang M., Maitituoheti M., Raman A.T., Ghosh A.K., Schulz J., Amin S.B., Orouji E., Tomczak K., Sarkar S., Oba J., Creasy C., Wu C.J., Khan S., Lazcano R., Wani K., Singh A., Barrodia P., Zhao D., Chen K., Haydu L.E., Wang W.L., Lazar A.J., Woodman S.E., Bernatchez C, & Rai K. (2021). Reprogramming of bivalent chromatin states in NRAS mutant melanoma suggests PRC2 inhibition as a therapeutic strategy. Cell reports, 36(3), 109410.
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8

Immunohistochemical Analysis of Aortic Tissue

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Formalin-fixed, paraffin embedded aortic tissue blocks were cut at 6μm thick. Slides were deparaffinized using Diva Decloaker (Biocare), blocked (Biocare, catalog No. RBM961) and stained for primary antibodies: Microsomal Glutathione S-Transferase 1 (MGST1; Abcam, catalog No. ab131059, 5.38μg/mL), ACTBL2 (ThermoFisher Scientific, catalog No. PA5-72402, 2μg/mL), CDH13 (Millipore, catalog No. abt121, 0.2μg/mL), LGALS9 (CellSignaling Technologies, catalog No. 55340, 0.27μg/mL), ORM2 (ThermoFisher Scientific, catalog No. PA5-37124, 5μg/mL), and PHB2 (CellSignaling Technology, catalog No. 14085, 1μg/mL). Secondary antibodies were used (Biocare, catalog No. RMR622) followed by 3,3′-Diaminobenzidine (DAB) chromogen and hematoxylin counterstain for nuclei. Slides were scanned and processed using QuPath (version 0.3.2, The University of Edinburgh) and images compiled in GraphPad Prism (version 8.0.0, San Diego, CA).
Tyrrell D.J., Chen J., Li B.Y., Wood S.C., Rosebury-Smith W., Remmer H.A., Jiang L., Zhang M., Salmon M., Ailawadi G., Yang B, & Goldstein D.R. (2022). Aging alters the aortic proteome in health and thoracic aortic aneurysm. Arteriosclerosis, thrombosis, and vascular biology, 42(8), 1060-1076.
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