Deep space black chromogen

Manufactured by Biocare Medical

Deep Space Black is a chromogen used in immunohistochemistry (IHC) and in situ hybridization (ISH) applications. It provides a dark black staining reaction when used with appropriate detection systems.

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Lab products found in correlation

Axioskop 2 microscope, by Zeiss (1 mentions) Bcl 2, by R&D Systems (1 mentions) Tissue tek oct media, by Sakura Finetek (1 mentions) Streptavidin hrp, by Biocare Medical (1 mentions) Vectra imaging system, by PerkinElmer (1 mentions) Warp red chromogen, by Biocare Medical (1 mentions) Inform 1, by PerkinElmer (1 mentions) Mach2 rabbit ap polymer, by Biocare Medical (1 mentions) Anti k8, by Developmental Studies Hybridoma Bank (1 mentions) Cat hematoxylin, by Biocare Medical (1 mentions) Bond polymer refine red detection kit, by Leica camera (1 mentions) Bond rx, by Leica camera (1 mentions) Bond polymer refine detection kit, by Leica camera (1 mentions) Rabbit anti iba1, by Fujifilm (1 mentions)

3 protocols using deep space black chromogen

Immunohistochemical Analysis of Kidney Tissues

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As vehicle mice became moribund, kidney tissues were collected and fixed in 10% neutral buffered formalin. In addition, after 24 weeks of treatment, representative mice from each group were euthanized. Kidneys were collected, bisected and either snap frozen in Tissue-Tek O.C.T. media (Sakura Finetek, Torrance, CA) or fixed in 10% neutral-buffered formalin and then paraffin-embedded.
Double- and single-label immunohistochemistry (IHC) was performed as essentially as described (13 (link)) with a primary antibody to BCL-2 (goat, R&D Systems, Minneapolis, MN), CD45R/B220 (rat, BD Biosciences, San Diego, CA), CD138 (rat, R&D Systems), CD3 (rabbit, Thermo Scientific) or IBA1 (rabbit, Wako, Japan), followed by secondary antibodies (Vector, Burlingame, CA), Leica Bond polymer for HRP or AP (Leica Biosystems, Buffalo Grove, IL), and Deep Space Black chromogen (Biocare Medical, Concord, CA). Following hematoxylin counterstaining, cells were imaged using a Zeiss Axioskop 2 microscope (Thornwood, NY).
IgG IF was performed with anti-mouse IgG Alexa 488 secondary antibody (chicken, Thermo Fisher Scientific) or IgY (IgG) -488 (chicken, Jackson ImmunoResearch) and semiquantitatively scored as previously described (23 (link)).

Ko K., Wang J., Perper S., Jiang Y., Yanez D., Kaverina N., Ai J., Liarski V.M., Chang A., Peng Y., Lan L., Westmoreland S., Olson L., Giger M.L., Wang L.C, & Clark M.R. (2016). BCL-2 as a Therapeutic Target in Human Tubulointerstitial Inflammation. Arthritis & rheumatology (Hoboken, N.J.), 68(11), 2740-2751.

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Immunohistochemical Analysis of Breast Cancer

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K5 and K8 expression were examined in a human breast cancer tissue microarray (TMA) containing patient tumors and normal breast samples (University of Wisconsin Carbone Cancer Center BioBank) (30 (link)). In brief, slides were deparaffinized, hydrated and permeabilized, then incubated with anti-K8 (Developmental Studies Hybridoma Bank, University of Iowa, TROMA-1), 1:50 in antibody diluent, (Da Vinci Green-BioCare Medical), followed by goat anti-rat IgG biotinylated secondary antibody (BioCare Medical), Streptavidin-HRP (BioCare Medical), and Deep Space Black chromogen (BioCare Medical). This was followed by anti-K5 (Covance Antibody Products, cat#PRB-160P), 1:4000, Mach 2 rabbit AP polymer (BioCare Medical) and Warp Red chromogen (BioCare Medical). Slides were counterstained with CAT hematoxylin (BioCare Medical), scanned using the Vectra imaging system (PerkinElmer Life Sciences), and analyzed using InForm 1.4 software (PerkinElmer Life Sciences), as described (30 (link)). Cytokeratin and hematoxylin signals were resolved using Nuance 3.0.2 (PerkinElmer Life Sciences), pseudocolored and merged to generate the images shown.

Shea M.P., O’Leary K.A., Fakhraldeen S.A., Goffin V., Friedl A., Wisinski K.B., Alexander C.M, & Schuler L.A. (2018). Anti-estrogen therapy increases plasticity and cancer stemness of prolactin-induced ERα+ mammary carcinomas. Cancer research, 78(7), 1672-1684.

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Immunohistochemical Analysis of Duodenal Tissues

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Duodenal segments were fixed in 10% buffered formalin and embedded in paraffin (FFPE) prior to sectioning and staining with H&E. 5 um FFPE sections were deparaffinized, rehydrated, and placed on a Leica Bond Rx immunohistochemical stainer for heat-activated antigen retrieval and sequential staining with Rabbit anti-human CD3 (Lab Vision), the Bond Polymer Refine Detection kit (Leica), and Deep Space Black Chromogen (Biocare Medical). Slides were then incubated with rabbit anti-IBA1 (Wako) and the Bond Polymer Refine Red Detection kit (Leica) and were counterstained with methyl-green. Isotype matched antibodies were used on serial tissue sections as negative controls.

Paustian A.M., Paez-Cortez J., Bryant S., Westmoreland S., Waegell W, & Kingsbury G. (2017). Continuous IL-23 stimulation drives ILC3 depletion in the upper GI tract and, in combination with TNFα, induces robust activation and a phenotypic switch of ILC3. PLoS ONE, 12(8), e0182841.

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