Discovery chromomap dab kit

Manufactured by Roche

Sourced in United States

The DISCOVERY ChromoMap DAB Kit is a laboratory equipment product designed for chromogenic detection and visualization of target antigens in immunohistochemistry (IHC) and in situ hybridization (ISH) applications. The kit provides the necessary reagents for a complete staining procedure, including the chromogenic substrate 3,3'-Diaminobenzidine (DAB) for the detection of target molecules.

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Lab products found in correlation

Ventana discovery ultra, by Roche (10 mentions) Hematoxylin, by Roche (5 mentions) Hematoxylin 2, by Roche (4 mentions) Bluing reagent, by Roche (4 mentions) Superfrost plus, by Thermo Fisher Scientific (3 mentions) Discovery ultra, by Roche (3 mentions) Cell conditioner 1, by Roche (3 mentions) Dmi8 microscope, by Leica (3 mentions) Discovery ultra antibody block, by Roche (3 mentions) Omni map anti rabbit hrp conjugated secondary antibody multimer hrp, by Roche (3 mentions) Anti phospho stat1 tyr701 58d6, by Cell Signaling Technology (3 mentions) Discovery omnimap anti rb hrp, by Roche (2 mentions) Ez prep solution, by Roche (2 mentions) Cell conditioning solution ultra cc1, by Roche (2 mentions) Discovery omnimap anti rt hrp, by Roche (2 mentions) Ventana discovery ultra autostainer, by Roche (2 mentions) Anti cd68 clone kp1, by Abcam (2 mentions) Anti hq hrp, by Roche (1 mentions) Anti rabbit hq, by Roche (1 mentions) Cell conditioning 1 cc1, by Roche (1 mentions)

Close spelling variants of this product

ChromoMap DAB kit (57 citations) ChromoMap DAB (23 citations) Discovery ChromoMap DAB (9 citations) ChromoMap kit (9 citations) Discovery ChromoMap DAB detection kit (8 citations) DAB kit (4 citations)

28 protocols using discovery chromomap dab kit

Automated Immunohistochemistry Staining for Abi1

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Immunohistochemistry was performed on TMA sections with the automated immunohistochemistry staining platform DISCOVERY ULTRA (Ventana Medical Systems, Inc.). Antigen retrieval was conducted with Cell Conditioning 1 (CC1) (Ventana) at 95 °C for 64 min. Slides were incubated with a 1:200 dilution of Abi1 antibody (Cell Signaling Technologies, 39444S) at room temperature for 2 h. For detection, a DISCOVERY ChromoMap DAB Kit, anti-HQ HRP, and anti-rabbit HQ (Ventana) were used. Digital images of stained TMAs were acquired with an SCN400 Slide Scanner (Leica Microsystems). Positively stained cells were analyzed with Aperio ImageScope (Leica Biosystems).

Nath D., Li X., Mondragon C., Post D., Chen M., White J.R., Hryniewicz-Jankowska A., Caza T., Kuznetsov V.A., Hehnly H., Jamaspishvili T., Berman D.M., Zhang F., Kung S.H., Fazli L., Gleave M.E., Bratslavsky G., Pandolfi P.P, & Kotula L. (2019). Abi1 loss drives prostate tumorigenesis through activation of EMT and non-canonical WNT signaling. Cell Communication and Signaling : CCS, 17, 120.

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Immunohistochemistry for Human CD3

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Tissues were fixed in 10% neutral buffered formalin, embedded in paraffin, sectioned at 4 µm, mounted on positive charged glass slides (Superfrost Plus; 12-550-15, Thermo Fisher) that were dried at 60°C for 20 min, and stained with H&E. The following immunohistochemistry protocol was used for the detection of human CD3: Anti-CD3 antibody (Clone EP41, Biocare Medical, CME324B), 60 min incubation, heat-induced epitope retrieval with cell conditioning media 1 (950-500; Ventana Medical Systems), 92 min; Visualization with DISCOVERY OmniMap anti-Rb HRP (760–4311; Ventana Medical Systems), DISCOVERY ChromoMap DAB kit (760-159; Ventana Medical Systems), Hematoxylin II (790–2208; Ventana Medical Systems), and Bluing reagent (790–2037; Ventana Medical Systems).

Beckett A.N., Chockley P., Pruett-Miller S.M., Nguyen P., Vogel P., Sheppard H., Krenciute G., Gottschalk S, & DeRenzo C. (2023). CD47 expression is critical for CAR T-cell survival in vivo. Journal for Immunotherapy of Cancer, 11(3), e005857.

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Immunohistochemical Analysis of 3D Cultures

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The 3D cultures were fixed in 10% neutral buffered formalin, paraffin embedded, sectioned using a microtome, and stained with H&E, according to commonly used procedures. The IHC stain was performed on a Ventana Discovery Ultra (Ventana Medical Systems, Roche Diagnostics, Indianapolis, IN, USA) automated IHC stainer. The slides were loaded onto the machine, and deparaffination, rehydration, endogenous peroxydase activity inhibition, and antigen retrieval were performed. Following primary antibody incubation, DISCOVERY anti-Rabbit HQ and DISCOVERY anti-HQ-HRP were incubated. The stains were then visualized with the DISCOVERY ChromoMap DAB Kit, counterstained with hematoxylin (Ventana). Primary antibody used: Ki-67: Clone# 30-9, rabbit monoclonal antibody; P53: Clone# DO-7, mouse monoclonal antibody (Ventana). Images were taken with a VENTANA iScan HT slide scanner and analyzed with a Ventana Image Viewer.

Chen M., Liang S., Shahid A., Andresen B.T, & Huang Y. (2020). The β-Blocker Carvedilol Prevented Ultraviolet-Mediated Damage of Murine Epidermal Cells and 3D Human Reconstructed Skin. International Journal of Molecular Sciences, 21(3), 798.

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Lung Tissue Histology Workflow

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Lung tissues were harvested and immediately fixed in 10% neutral buffered formalin. Dehydration, clear, and paraffinization were performed on a Tissue‐Tek VIP Vacuum Infiltration Processor (SAKURA). The samples were embedded in paraffin using a Tissue‐Tek TEC Tissue Embedding Station (SAKURA). Samples were then sectioned at 5 μm and put on positively charged glass slides. The slides were deparaffinized, rehydrated, and stained with Modified Mayer's hematoxylin and Eosin Y (H&E) Stain (America MasterTech Scientific) on an H&E Auto Stainer (Prisma Plus Auto Stainer, SAKURA) according to standard laboratory procedures. The stains were visualized with DISCOVERY ChromoMap DAB Kit, counterstained with hematoxylin (Ventana) and cover‐slipped. H&E stained slides were digitalized and documented by iScan HT (Roche) scanner, and then representative pictures (10× magnified) were taken from the NDP.view2 viewing software.

Williams L., Li L., Yazaki P.J., Wong P., Hong T., Poku E.K., Hui S., Ghimire H., Shively J.E, & Kujawski M. (2024). Comparison of IL‐2‐antibody to IL‐2‐Fc with or without stereotactic radiation therapy in CEA immunocompetent mice with CEA positive tumors. Cancer Medicine, 13(3), e6909.

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Quantifying Capillary Density in Ischemic Tissues

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To analyze the capillary density in treated and untreated ischemic tissues, immunostaining of muscle slices was performed at day 21 post treatment. For that, samples were treated with Cell Conditioning 1 buffer (CC1) TRIS pH 8 during 32 min before incubation with the primary antibody. Subsequently, the primary anti–CD31 monoclonal antibody (ab182981, Abcam) was added at 1:200 dilution and incubated for 40 min at room temperature. OmniMap Discovery anti-Rb HRP (Ventana Medical Systems, Roche) was used as secondary antibody. Finally, the signal was detected using the Discovery ChromoMap DAB kit (Ventana Medical Systems, Roche). Vascularization was determined by the density of CD31 positively stained vessels, quantified as the number of capillaries per muscle fiber in ten fields per muscle. Four mice were analyzed for each condition.

Ibáñez-Fonseca A., Rico A., Preciado S., González-Pérez F., Muntión S., García-Briñón J., García-Macías M.C., Rodríguez-Cabello J.C., Pericacho M., Alonso M, & Sánchez-Guijo F. (2022). Mesenchymal Stromal Cells Combined With Elastin-Like Recombinamers Increase Angiogenesis In Vivo After Hindlimb Ischemia. Frontiers in Bioengineering and Biotechnology, 10, 918602.

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Quantitative IHC Analysis of Breast Carcinoma

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Immunohistochemistry (IHC) analysis of primary and matched metastatic breast carcinoma tissue array (BR10010e from US Biomax) was performed by US Biomax. Briefly, slides were stained on a Ventana Discovery Ultra automated system, using the manufacturer’s protocol with proprietary reagents. Heat-induced antigen retrieval (HIER) was performed with CC1, pH 8.5 (Ventana) for 56 minutes at 95 °C. Peroxidase inhibitor was applied. The primary antibody (CHAF1B-HPA021679, Sigma-Aldrich at 1:50) was incubated at 37 °C for 28 minutes. The secondary an tibody used was Discovery anti-Rabbit HQ and anti-HQ HRP (Ventana), incubated at 37 °C for 8 minutes each. Tyramide signal amplification was applied using Discovery Amp HQ kit (Ventana), for 8 minutes. DAB chromagen was applied using Discovery ChromoMap DAB Kit (Ventana). Slides were then counterstained with Hematoxylin II (Ventana). Images were quantified using Visiopharm, an automated quantification software. Nuclei were scored as Negative, 1+, 2+, and 3+ using the algorithm for ER/PR scoring. For each sample an H-score (Sum of percentages of 1+ 2+ 3+ nuclei weighted by a factor of 1, 2, and 3 respectively) was calculated.

Gomes A.P., Ilter D., Low V., Rosenzweig A., Shen Z.J., Schild T., Rivas M.A., Er E.E., McNally D.R., Mutvei A.P., Han J., Ou Y.H., Cavaliere P., Mullarky E., Nagiec M., Shin S., Yoon S.O., Dephoure N., Massagué J., Melnick A.M., Cantley L.C., Tyler J.K, & Blenis J. (2019). Dynamic incorporation of histone H3 variants into chromatin is essential for acquisition of aggressive traits and metastatic colonization. Cancer cell, 36(4), 402-417.e13.

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Immunohistochemical Detection of pAkt in Brain Sections

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The 4-μm brain sections were deparaffinized in an automated system (Discovery XT; Ventana Medical Systems) with EZ Prep solution. A heat-induced antigen retrieval method was used in Cell Conditioning 1 solution. Rabbit primary antibody reacting with pAkt (ab81283; Abcam, Cambridge, MA) was used at a 1:200 dilution in PSS Diluent for antibodies (Ventana Medical Systems) and was incubated for 32 min, followed by OmniMap anti-rabbit secondary antibody (Ventana Medical Systems) for 20 min. For detection, the Discovery ChromoMap DAB Kit (Ventana Medical Systems) was used; slides were then counterstained with hematoxylin, dehydrated, and coverslipped. A BX53 Microscope (Olympus) and cellSens Dimension software were used to obtain pictures with ×10 and ×60 objectives.

Sajan M., Hansen B., Ivey R I.I.I., Sajan J., Ari C., Song S., Braun U., Leitges M., Farese-Higgs M, & Farese R.V. (2016). Brain Insulin Signaling Is Increased in Insulin-Resistant States and Decreases in FOXOs and PGC-1α and Increases in Aβ1–40/42 and Phospho-Tau May Abet Alzheimer Development. Diabetes, 65(7), 1892-1903.

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Immunohistochemical Staining of Mouse Tumor Tissues

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For immunohistochemistry (IHC) staining, nude mouse tumor tissues were fixed in 10% neutral formalin (NBF), followed by paraffin embedding. Then the tumor block was cut into 5 μm thick slices and placed on positively charged slides. IHC staining was performed on the Ventana Discovery Ultra (Ventana Medical Systems, Roche Diagnostics, Indianapolis, USA) IHC automatic staining machine. The slides were loaded on the machine and subjected to deparaffinization, rehydration, endogenous peroxidase activity inhibition, and antigen retrieval. Then, the slides were incubated with Ki67 antibody (ab16667, Abcam, UK). IHC tumor sections were visualized using the DISCOVERY-ChromoMap-DAB kit (Ventana), and Suomijin (Ventana) was used for counterstaining [31] . The slides were imaged using the Leica Dmi8 microscope and analyzed using image-pro Premier software.

Huang G., Wu Y., Gan H, & Chu L. (2023). Overexpression of CD2/CD27 could inhibit the activation of nitrogen metabolism pathways and suppress M2 polarization of macrophages, thereby preventing brain metastasis of breast cancer. Translational Oncology, 37, 101768.

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Histological Analysis of SARS-CoV-2 Receptor Expression

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Tissues were fixed in 10% neutral buffered formalin for a minimum of seven days. Sagittal-cut skulls were decalcified in 20% EDTA in sucrose (Newcomer Supply, Middleton, WI, USA), changed weekly for six weeks. Tissues were then processed using a Sakura VIP-6 Tissue Tek tissue processor and embedded in Ultraffin paraffin polymer (Cancer Diagnostics, Durham, NC, USA). Samples were sectioned at 5 µm, and resulting slides were stained with hematoxylin and eosin. For WIV1-CoV immunohistochemistry (IHC), tissues were stained with SARS NP rabbit polyclonal antibody (Novus Biologicals, NB100-56576, 1:250, Centennial, CO, USA). For ACE2 IHC, tissues were stained with ACE2 rabbit polyclonal antibody (Abcam, ab15348, 1:1500). Hereafter, antibodies were detected using ImmPress VR Polymer HRP anti-rabbit IgG (Vector Laboratories, MP-6401-15, no dilution, Burlingame, CA, USA) followed by Discovery ChromoMap DAB kit (Ventana Medical Systems, 760-159, Tucson, AZ, USA).

van Doremalen N., Schäfer A., Menachery V.D., Letko M., Bushmaker T., Fischer R.J., Figueroa D.M., Hanley P.W., Saturday G., Baric R.S, & Munster V.J. (2018). SARS-Like Coronavirus WIV1-CoV Does Not Replicate in Egyptian Fruit Bats (Rousettus aegyptiacus). Viruses, 10(12), 727.

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Immunohistochemical Analysis of Ki67 Expression

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Tumor tissues were fixed in 10% neutral formalin, embedded in paraffin and cut into 5-μm-thick sections. Immunohistochemistry was performed on a Ventana Discovery Ultra automatic dyeing machine (Ventana Medical Systems, Roche Diagnostics, Indianapolis). The cover glass was loaded on the machine whereupon the sections were dewaxed and rehydrated, followed by endogenous peroxidase activity elimination and antigen retrieval. The sections were incubated with anti-Ki67 antibody (ab16667, Abcam), developed with DISCOVERY ChromoMap DAB Kit and counterstained with hematoxylin (Ventana). The images were collected with a DMi8 Leica microscope and analyzed with Image-Pro Premier software.

Chi Y., Xin H, & Liu Z. (2021). Exosomal lncRNA UCA1 Derived From Pancreatic Stellate Cells Promotes Gemcitabine Resistance in Pancreatic Cancer via the SOCS3/EZH2 Axis. Frontiers in Oncology, 11, 671082.

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