Vina green chromogen

Manufactured by Biocare Medical

Vina Green Chromogen is a laboratory reagent used in histochemical and immunohistochemical staining procedures. It is a substrate that undergoes an enzymatic reaction, resulting in the formation of a green colored product. The core function of Vina Green Chromogen is to provide a visible signal that indicates the presence of a target analyte in a sample.

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Vina Green (11 citations) Vina Green Chromogen Kit (8 citations)

6 protocols using vina green chromogen

Immunohistochemical Detection of Salmonella

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Paraffin-embedded tissue samples were sectioned at
a thickness of
3–4 μm on charged microscope slides. Samples were deparaffinized
using CitriSolv (Decon Laboratories, Inc., King of Prussia, PA) and
rehydrated sequentially in graded alcohols. Antigen retrieval was
performed by incubating slides in 10 μg/mL proteinase K (MilliporeSigma)
in PK buffer (0.6 M Tris (pH 7.5)/0.1% CaCl₂) for 10 min
at RT. Blocking of endogenous peroxidase and alkaline phosphatase
was performed by incubating slides in Rodent Block M (BioCare Medical,
Pacheco, CA) followed by incubation with BLOXALL Endogenous Blocking
Solution (Vector Laboratories, Burlingame, CA). Slides were washed
in TBS buffer and incubated with primary rabbit Salmonella O Antiserum
(Group B Factors 1, 4, 5, 12, #BD 229481, Becton, Dickinson and Company)
for 1 h at 1:5000 dilution. Slides were then incubated in AP-polymer
(Rabbit on Rodent AP-Polymer, BioCare Medical) followed by Vina Green
Chromogen (BioCare Medical, Pacheco, CA) treatment. Tissues were finally
counterstained with hematoxylin (BioCare Medical) before mounting
with EcoMount (BioCare Medical). Slides were cured at 60–70
°C for 15 min.

Richards A.F., Baranova D.E., Pizzuto M.S., Jaconi S., Willsey G.G., Torres-Velez F.J., Doering J.E., Benigni F., Corti D, & Mantis N.J. (2021). Recombinant Human Secretory IgA Induces Salmonella Typhimurium Agglutination and Limits Bacterial Invasion into Gut-Associated Lymphoid Tissues. ACS Infectious Diseases, 7(5), 1221-1235.

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Multimodal Immunohistochemical Staining Protocol for Parasite Identification

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After the HIER, the slides were treated with 3% hydrogen peroxide solution for 15 minutes to block endogenous peroxidase activity, followed by blocking with TBST with BSA. The slides were then incubated with goat anti-GFAP, goat anti-Iba1, or rabbit anti-NeuN antibodies (Abcam, Cambridge, MA) overnight at 4°C. The slides were washed in TBST and incubated with horseradish peroxidase (HRP)-conjugated donkey anti-goat IgG antibody (Invitrogen, Carlsbad, CA) or goat anti-rabbit IgG antibody (Invitrogen) for one hour at room temperature. Color was developed using diaminobenzidine (Vector Laboratories, Burlingame, CA). Thereafter, the slides were subjected to one more round of HIER to facilitate removal of the pre-bound antibodies used for the staining for the cell markers [21 (link)]. After re-blocking, the slides were then incubated with polyclonal rabbit anti-T. gondii serum [20 (link)] for one hours at room temperature, and then after washing, incubated with the HRP-conjugated goat anti-rabbit IgG antibody. Thereafter, the slides were incubated with Vina Green Chromogen (Biocare Medical) for color development. The slides were examined using Nikon Eclipse 90i microscope, and images were obtained using a digital camera and NIS elements BR 3.2 software (Nikon).

Anand N., Lutshumba J., Whitlow M., Abdelaziz M.H., Mani R, & Suzuki Y. (2021). Deficiency in indoleamine-2, 3-dioxygenase induces upregulation of guanylate binding protein 1 and inducible nitric oxide synthase expression in the brain during cerebral infection with Toxoplasma gondii in genetically resistant BALB/c mice but not in genetically susceptible C57BL/6 mice. Microbes and infection, 24(3), 104908.

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Multiplex Immunohistochemistry for Foxp3 and CD3

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Sections were fixed in acidic methanol (60% methanol, 10% acetic acid, 30% water), embedded in paraffin and sectioned at 5 micron thickness. Foxp3 and CD3 T cells were demonstrated by sequential double-staining method using rat monoclonal antibody to Foxp3 (eBioscience, clone: FJK-16s) and Rabbit polyclonal antibody to CD3 (Abcam Inc) after heat induced antigen retrieval. As secondary antibodies Rat HRP-Polymer (Biocare Medical) used for Foxp3 and Rabbit AP-Polymer (Biocare Medical) for CD3. Foxp3⁺ cells were stained with Vina Green chromogen (Biocare Medical) and CD3⁺ cells with Vulcan Fast Red (Biocare Medical). Hematoxylin was used for nuclear counterstain.

Do J.S., Visperas A., Sanogo Y.O., Bechtel J.J., Dvorina N., Kim S., Jang E., Stohlman S.A., Shen B., Fairchild R.L., Baldwin WM I.I.I., Vignali D.A, & Min B. (2015). An IL-27/Lag3 axis enhances Foxp3+ regulatory T cell suppressive function and therapeutic efficacy. Mucosal immunology, 9(1), 137-145.

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Multiplex Immunohistochemistry for Foxp3 and CD3

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Antimicrobial peptide expression in human skin

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Infected human skin explants treated with Aquaphor or 2% PALA were fixed in Histochoice. Next, paraffin embedded samples were sectioned and stained for HBD2, HBD3, or LL-37. Briefly, dewaxed and rehydrated sections were first subjected to antigen retrieval in sodium citrate buffer using a steamer for 25 minutes. Primary antibody staining was performed at 4 °C overnight in a humidifying chamber. Labelled polymer-HRP anti-rabbit (Dako, K4011), biotinylated secondary goat anti-mouse (EMD Millipore, IHC Select, 20775), and streptavidin-HRP (EMD Millipore, IHC Select, 20774) were used as detection reagents/secondary antibodies. Vina green chromogen (Biocare Medical, BRR807A) was used to visualize protein levels. Aperio AT2 (Leica Biosystems) was used to scan the slides.

Jatana S., Homer C.R., Madajka M., Ponti A.K., Kabi A., Papay F, & McDonald C. (2018). Pyrimidine synthesis inhibition enhances cutaneous defenses against antibiotic resistant bacteria through activation of NOD2 signaling. Scientific Reports, 8, 8708.

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Dual Immunohistochemical Staining Protocol

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After tissue slides were prepared, they were incubated with the first antibody goat anti-rabbit HRP polymer (Mach 2 Rabbit HRP-Polymer, Biocare Medical), and Betazoid DAB Chromogen (Biocare Medical) was then used to stain the first marker. The slides were rinsed thoroughly with distilled water, treated with denaturing solution (Biocare Medical) for 3 min to denature anti-rabbit-HRP, and then rinsed for staining with the second antibody. Goat anti-mouse HRP polymer (Mach 2 Mouse HRP-Polymer, Biocare Medical) and Vina Green Chromogen (Biocare Medical) were used for the staining. The slides were finally counterstained with hematoxylin (Biocare Medical).

Wang Q., Jiang J., Ying G., Xie X.Q., Zhang X., Xu W., Zhang X., Song E., Bu H., Ping Y.F., Yao X.H., Wang B., Xu S., Yan Z.X., Tai Y., Hu B., Qi X., Wang Y.X., He Z.C., Wang Y., Wang J.M., Cui Y.H., Chen F., Meng K., Wang Z, & Bian X.W. (2018). Tamoxifen enhances stemness and promotes metastasis of ERα36+ breast cancer by upregulating ALDH1A1 in cancer cells. Cell Research, 28(3), 336-358.

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