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Betazoid dab chromagen kit

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The Betazoid DAB Chromagen Kit is a laboratory reagent used for the detection and visualization of targeted biomolecules in biological samples during immunohistochemical and in situ hybridization procedures. The kit utilizes 3,3'-Diaminobenzidine (DAB) as the chromogenic substrate to produce a brown reaction product at the site of the target analyte.

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Lab products found in correlation

Normal goat serum (ngs), by Vector Laboratories (3 mentions) Brucella polyclonal rabbit antibody, by Bioss Antibodies (3 mentions) Bloxall blocking solution, by Vector Laboratories (3 mentions) Ems solution a, by Electron Microscopy Sciences (2 mentions) Vectastain abc, by Biocare Medical (2 mentions) Mach4 universal hrp polymer kit, by Biocare Medical (1 mentions) Peroxidazed 1, by Biocare Medical (1 mentions) Background sniper, by Biocare Medical (1 mentions) Rabbit on rodent hrp polymer, by Biocare Medical (1 mentions) Cleaved caspase 3 primary antibody, by Cell Signaling Technology (1 mentions) Vectastain elite abc hrp kit, by Vector Laboratories (1 mentions)

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Betazoid DAB (15 citations) DAB chromagen (3 citations)

5 protocols using betazoid dab chromagen kit

1

Immunohistochemical Detection of Brucella in Tissues

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Unstained slides from liver, spleen, uterus, and mesenteric lymph nodes were adhered to positively charged glass slides for immunohistochemistry. Slides were deparaffinized and rehydrated through a series of xylene and ethanol steps before antigen retrieval was performed using 1:10 EMS Solution A (Electron Microscopy Services) in a 2100 Antigen Retriever (Aptum Biologics Ltd.), according to the manufacturer’s instruction. Endogenous peroxidase activity was blocked by 10 m incubation with Bloxall Blocking Solution (Vector Laboratories) followed by 20 m blocking with normal goat serum (Vector). After each step slides were washed with PBS plus 0.5% tween for 5 minutes. Primary incubation was overnight at 4°C with Brucella polyclonal rabbit antibody (Bioss) at 1:400. Negative control tissues were incubated with rabbit nonimmune serum diluted in PBS. A Vectastain ABC and Betazoid DAB chromagen kits (Biocare Medical) were used following primary incubation according to the manufacturer’s instructions. The slides were counterstained with Meyer’s hematoxylin III.
Stranahan L.W., Khalaf O.H., Garcia-Gonzalez D.G, & Arenas-Gamboa A.M. (2019). Characterization of Brucella canis infection in mice. PLoS ONE, 14(6), e0218809.
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2

Immunohistochemical Detection of Brucella in Tissues

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Unstained slides from spleen, uterus, liver, and lung were adhered to positively charged glass slides for immunohistochemistry. Slides were deparaffinized and rehydrated through a series of xylene and ethanol steps before antigen retrieval was performed using 1:10 EMS Solution A (Electron Microscopy Services) in a 2100 Antigen Retriever (Aptum Biologics Ltd.), according to manufacturer protocol. Endogenous peroxidases were blocked by 10 m incubation with Bloxall Blocking Solution (Vector Laboratories) followed by 20 m blocking with normal goat serum (Vector). After each step slides were washed with PBS plus 0.5% tween for 5 minutes. Primary incubation was overnight at 4o C with Brucella polyclonal rabbit antibody (Bioss) at 1:600. Negative control tissues were incubated with rabbit nonimmune serum diluted in PBS. A Vectastain ABC and Betazoid DAB chromagen kits (Biocare Medical) were used following primary incubation according to the manufacturer’s instructions. The slides were counterstained with Meyer’s hematoxylin III.
Hensel M.E., Garcia-Gonzalez D.G., Chaki S.P., Samuel J, & Arenas-Gamboa A.M. (2019). Characterization of an intratracheal aerosol challenge model of Brucella melitensis in guinea pigs. PLoS ONE, 14(3), e0212457.
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3

SARS-CoV-2 N Protein Immunostaining Protocol

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Immunostainings were performed as previously described (Chen et al., 2020 (link)). Briefly, 5 um lung tissues sections were mounted on Superfrost Plus Microscope slides, deparaffinized and antigen retrieval was conducted using the HIER method (Heat Induced Epitope Retrieval). Subsequently, slides were stained with the primary N protein monoclonal Ab 1C7 (1 μg/ml). Then slides were incubated and developed with the MACH 4 Universal HRP Polymer Kit and Betazoid DAB Chromagen Kit (Biocare Medical, LLC), according to the manufacturer’s instructions. The presence of SARS-CoV-2 N protein antigen was analysed by a veterinary pathologist at Texas Biomedical Research Institute. Scores of antigens were graded based on the anatomical location of viral N protein staining: grade 0 = no or rare immunostaining, grade 1 = viral N protein staining only in bronchi/bronchiole, grade 2 = viral N protein staining only in bronchi/bronchiole and surrounding alveolar septa, grade 3 = viral N protein staining in alveolar septa distant from the bronchi and bronchioles, grade 4 = viral N protein staining throughout the lung. The Mann–Whitney test was used for statistical analysis.
Park J.G., Oladunni F.S., Rohaim M.A., Whittingham-Dowd J., Tollitt J., Hodges M.D., Fathallah N., Assas M.B., Alhazmi W., Almilaibary A., Iqbal M., Chang P., Escalona R., Shivanna V., Torrelles J.B., Worthington J.J., Jackson-Jones L.H., Martinez-Sobrido L, & Munir M. (2021). Immunogenicity and protective efficacy of an intranasal live-attenuated vaccine against SARS-CoV-2. iScience, 24(9), 102941.
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4

Quantification of Apoptotic Hepatocytes

Cited 1 time
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Formalin-fixed tissue sections (4 μm) from 4 males and 4 females from each treatment group (24h timepoint, same mice as those analyzed for γH2AX and histopathology) were deparaffinized in xylenes, rehydrated, and subjected to HIER in Buffer H, pH 8.8 (Thermo Scientific). After cooling, endogenous peroxidase was blocked with Peroxidazed 1 (Biocare Medical) for 5 minutes and slides were blocked in Background Sniper (Biocare Medical) for 15 minutes. Slides were then incubated with cleaved caspase-3 primary antibody (Cell Signaling Technology) for 60 minutes at room temperature, washed, and incubated with Rabbit-on-Rodent HRP Polymer (Biocare Medical) for 30 minutes at room temperature. Slides were washed and incubated with DAB (Betazoid DAB Chromagen kit, Biocare Medical), washed, and counterstained with hematoxylin. Stained slides were scanned with a Leica Aperio AT2 slide scanning microscope and analyzed with QuPath software (Bankhead et al., 2017 (link)). After blinding filenames, 4 equal-sized, randomly selected, independent regions of tissue were annotated for number of nuclei and number of apoptotic events. The percentage of apoptotic events for each animal was calculated from the total number of apoptotic hepatocytes and total number of nuclei from four representative 400x fields.
Kay J.E., Corrigan J.J., Armijo A.L., Nazari I.S., Kohale I.N., Torous D.K., Avlasevich S.L., Croy R.G., Wadduwage D.N., Carrasco S.E., Dertinger S.D., White F.M., Essigmann J.M., Samson L.D, & Engelward B.P. (2021). Excision of mutagenic replication-blocking lesions suppresses cancer but promotes cytotoxicity and lethality in nitrosamine-exposed mice. Cell reports, 34(11), 108864.
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5

Immunohistochemical Detection of Brucella

Cited 1 time
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Five micrometer tissue sections of testicle/epididymis (mouse and guinea pig) and prostate (guinea pig only) were adhered to positively charged glass slides for immunohistochemistry. Slides were routinely processed, and antigen retrieval was performed as previously described using a 2,100 Antigen Retriever (Aptum Biologics Ltd. Southampton, UK; Hensel et al., 2019 (link)). Slides were blocked as previously described with Bloxall Blocking Solution (Vector Laboratories, Burlingame, CA) and normal goat serum (Vector Laboratories; Hensel et al., 2019 (link)). Primary incubation was performed overnight at 4o C with a Brucella polyclonal rabbit antibody (Bioss Antibodies, Boston, MA) at dilution of 1:500. A Vectastain Elite® ABC HRP Kit (Vector Laboratories) with an avidin/biotinylated anti-rabbit secondary antibody was used according to the manufacturer’s instructions. Antigen was visualized with a Betazoid DAB chromagen kit (Biocare Medical, Pachecho, CA). The slides were counterstained with Gills’s hematoxylin III and cover slipped.
Hensel M.E., Stranahan L.W., Edwards J.F, & Arenas-Gamboa A.M. (2022). Intratracheal inoculation results in Brucella-associated reproductive disease in male mouse and guinea pig models of infection. Frontiers in Microbiology, 13, 1029199.
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