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Mach3 mouse hrp polymer detection

Manufactured by Biocare Medical
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The MACH3 Mouse HRP-Polymer Detection is a laboratory equipment product designed for immunohistochemistry applications. It is a polymer-based detection system that utilizes horseradish peroxidase (HRP) as the enzymatic label. The core function of this product is to provide a sensitive and reliable method for the visualization of target antigens in tissue sections or cell preparations.

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Lab products found in correlation

Background sniper, by Biocare Medical (2 mentions) H550s microscope, by Nikon (2 mentions) Cd68 kp1, by Agilent Technologies (2 mentions) Diva decloaker, by Biocare Medical (2 mentions) Cd45 lca 2b11 pd7 26, by Agilent Technologies (2 mentions) Cd3 sp7, by Thermo Fisher Scientific (2 mentions) Cd20 l26, by Biocare Medical (2 mentions) Opal 570, by Akoya Biosciences (1 mentions) Axioplan 2, by Leica (1 mentions) Rm2245 microtome, by Leica (1 mentions) Immpress anti rabbit igg polymer kit, by Vector Laboratories (1 mentions)

4 protocols using mach3 mouse hrp polymer detection

1

Immunohistochemical Analysis of Human Cells

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Tissues for IHC were harvested from MoM and fixed in 4% paraformaldehyde for 24 hour at 4°C, embedded in paraffin, cut into 5-μm sections and mounted onto poly-L-lysine coated glass slides. Following paraffin removal, antigen retrieval (DIVA Decloaker, Biocare Medical, Cat. DV2004.) and blocking of non-specific Ig-binding sites (Background Sniper, Biocare Medical), tissue sections were stained with primary antibodies overnight at 4°C and developed with a biotin-free horseradish peroxidase (HRP)-polymer system (MACH3 Mouse HRP-Polymer Detection, Biocare Medical). All tissue sections were then counterstained with hematoxylin. Primary antibodies directed against CD45 LCA (2B11&PD7/26, Dako), CD3 (SP7, Thermo Scientific), CD20 (L26, Biocare Medical) and CD68 (KP1, Dako) were used to identify human cells in the spleen, liver and lung. For comparison, tissue sections were stained with mouse IgG1k or IgG2a isotype controls. Light microscopy images were taken with a Nikon H550S microscope at 40×.
Honeycutt J.B., Thayer W.O., Baker C.E., Ribeiro R.M., Lada S.M., Cao Y., Cleary R.A., Hudgens M.G., Richman D.D, & Garcia J.V. (2017). HIV persistence in tissue macrophages of humanized myeloid only mice during antiretroviral therapy. Nature medicine, 23(5), 638-643.
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2

Immunohistochemical Analysis of Human Cells

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Tissues for IHC were harvested from MoM and fixed in 4% paraformaldehyde for 24 hour at 4°C, embedded in paraffin, cut into 5-μm sections and mounted onto poly-L-lysine coated glass slides. Following paraffin removal, antigen retrieval (DIVA Decloaker, Biocare Medical, Cat. DV2004.) and blocking of non-specific Ig-binding sites (Background Sniper, Biocare Medical), tissue sections were stained with primary antibodies overnight at 4°C and developed with a biotin-free horseradish peroxidase (HRP)-polymer system (MACH3 Mouse HRP-Polymer Detection, Biocare Medical). All tissue sections were then counterstained with hematoxylin. Primary antibodies directed against CD45 LCA (2B11&PD7/26, Dako), CD3 (SP7, Thermo Scientific), CD20 (L26, Biocare Medical) and CD68 (KP1, Dako) were used to identify human cells in the spleen, liver and lung. For comparison, tissue sections were stained with mouse IgG1k or IgG2a isotype controls. Light microscopy images were taken with a Nikon H550S microscope at 40×.
Honeycutt J.B., Thayer W.O., Baker C.E., Ribeiro R.M., Lada S.M., Cao Y., Cleary R.A., Hudgens M.G., Richman D.D, & Garcia J.V. (2017). HIV persistence in tissue macrophages of humanized myeloid only mice during antiretroviral therapy. Nature medicine, 23(5), 638-643.
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3

Validating SARS-CoV-2 Spike Protein Detection

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Antibodies and probes for detecting SARS-CoV-2 spike protein (s-SARS-CoV-2-s protein) in formalin-fixed paraffin-embedded (FFPE) tissue were first validated on SARS-CoV-2 infected and non-infected CACO2 cells (ATCC, HTB-37) that were processed to FFPE blocks (supplementary Fig. 1). For immunohistochemistry of s-SARS-CoV-2-s protein, the SARS-CoV-2-s antibody (clone 1A9, Genetex, USA) was used at a dilution of 1:80. Detection was carried out with MACH 3 Mouse HRP Polymer detection (Biocare Medical, USA) following the manufacturer´s protocol. For in situ hybridization of SARS-CoV-2 a sense probe (RNAscope CoV2019-S-sense, ACD Bio-Techne, USA) and an antisense probe (RNAscope CoV2019-S-antisense, ACD Bio-Techne) was used. Detection was carried out with Opal 570 (Akoya Bioscience) according to the manufacturer´s protocol. All slides were evaluated independently by two experienced pathologists (MR, HB).
Paul T., Ledderose S., Bartsch H., Sun N., Soliman S., Märkl B., Ruf V., Herms J., Stern M., Keppler O.T., Delbridge C., Müller S., Piontek G., Kimoto Y.S., Schreiber F., Williams T.A., Neumann J., Knösel T., Schulz H., Spallek R., Graw M., Kirchner T., Walch A, & Rudelius M. (2022). Adrenal tropism of SARS-CoV-2 and adrenal findings in a post-mortem case series of patients with severe fatal COVID-19. Nature Communications, 13, 1589.
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4

Histological Analysis of Organoid Sections

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Organoids were fixed with 4% PFA for 1 h at RT, washed with PBS and embedded in paraffin. Organoid sections of 1.5 µm thickness were generated using the Leica RM2245 microtome (Leica). Organoid sections were stained with H/E or Gomori using standard protocols or used for immunohistochemistry staining. After antigen retrieval, slides were incubated with CD34 (Sigma-Aldrich, catalog no. QEBnd-10; 1:100) or MPO (DAKO/Agilent, catalog no. A0298, 1:400) for 1 h. Detection was carried out with ImmPress anti-rabbit IgG polymer kit (Vector Laboratories) or MACH 3 Mouse HRP Polymer detection (Biocare Medical), respectively, according to the manufacturer´s protocol. Images were acquired on a Carl Zeiss Axioplan 2 or a Leica DM 2000 microscope. Reticulin staining was assessed by an experienced hematopathologist.
Frenz-Wiessner S., Fairley S.D., Buser M., Goek I., Salewskij K., Jonsson G., Illig D., zu Putlitz B., Petersheim D., Li Y., Chen P.H., Kalauz M., Conca R., Sterr M., Geuder J., Mizoguchi Y., Megens R.T., Linder M.I., Kotlarz D., Rudelius M., Penninger J.M., Marr C, & Klein C. (2024). Generation of complex bone marrow organoids from human induced pluripotent stem cells. Nature Methods, 21(5), 868-881.
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