Vina green chromogen kit

Manufactured by Biocare Medical

Sourced in United States, Japan

The Vina Green Chromogen Kit is a laboratory reagent used for the detection and visualization of specific molecular targets in biological samples. The kit provides a green-colored chromogenic substrate that reacts with enzymes, such as horseradish peroxidase (HRP) or alkaline phosphatase (AP), to produce a visible signal. This signal can be used to identify the presence and location of the target molecules within the sample.

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Lab products found in correlation

Rabbit anti mouse cd8α d4w2z, by Cell Signaling Technology (2 mentions) Anti mouse cxcl5, by LSBio (2 mentions) Nexgen decloaker chamber, by Biocare Medical (2 mentions) Warp red chromagen kit, by Biocare Medical (2 mentions) Rabbit anti mouse β catenin, by Cell Signaling Technology (2 mentions) Rat anti mouse ly6g, by Abcam (2 mentions) Clsm 700 confocal microscope, by Zeiss (2 mentions) Rabbit goat mouse abc kit, by Vector Laboratories (1 mentions) Histofine dab substrate, by Nichirei Biosciences (1 mentions) Horseradish peroxidase, by Agilent Technologies (1 mentions) Denaturing solution, by Biocare Medical (1 mentions) Dextran polymers, by Agilent Technologies (1 mentions) Wash buffer, by Agilent Technologies (1 mentions) Phospho stat1, by Cell Signaling Technology (1 mentions) Envision system, by Agilent Technologies (1 mentions) Histofine simple stain dab, by Nichirei Biosciences (1 mentions) Imagescope, by Leica Microsystems (1 mentions) Immpact vector red ap substrate kit, by Vector Laboratories (1 mentions) Bz x700, by Keyence (1 mentions) Peroxide block, by ScyTek Laboratories (1 mentions)

6 protocols using vina green chromogen kit

Immunohistochemical Analysis of SHH and PTCH

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IHC was carried out using the antibodies detailed in Table 1. Following antigen retrieval, sections were treated with 10% normal serum for 30 min, and then incubated with primary antibodies at 4 °C overnight. Tagging of primary antibody was achieved by the subsequent application of anti-rabbit, anti-goat, or anti-mouse IgG and avidin–biotin complexes (Rabbit/Goat/Mouse ABC kit; Vector Laboratories, Inc., Burlingame, CA, USA). Immunoreactivity was visualized using diaminobenzidine (DAB)/H₂O₂ solution (Histofine DAB substrate; Nichirei, Tokyo, Japan), and sections were counterstained with Mayer’s hematoxylin. For double immunostaining, the same process from DAB staining was repeated, and immunoreactivity was visualized using Green chromogen (Vina Green Chromogen kit; Biocare Medical, Pacheco, CA, USA). Sections were counterstained with Mayer’s hematoxylin.
SHH and PTCH expression in OSCC were evaluated independently by the 2 pathologists. The results were scored from 0 to 3 based on the intensity of the staining at the membrane or in the cytoplasm: 0, no reactivity; +1, weak; +2, moderate; and 3, marked.

Takabatake K., Shimo T., Murakami J., Anqi C., Kawai H., Yoshida S., Wathone Oo M., Haruka O., Sukegawa S., Tsujigiwa H., Nakano K, & Nagatsuka H. (2019). The Role of Sonic Hedgehog Signaling in the Tumor Microenvironment of Oral Squamous Cell Carcinoma. International Journal of Molecular Sciences, 20(22), 5779.

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Multicolor Immunohistochemistry Analysis

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Blocks were sectioned at a thickness of 3 μm. Antibodies targeting CD68 (clone PG-M1, Dako), phosphorylated signal transducer and activator of transcription 1 (phospho-STAT1; monoclonal, clone, and 58D6, Cell Signaling Technology, Danvers, MA, USA), and c-Maf (clone EPR16484, Abcam, Cambridge, UK) were used for analyses. Double staining was performed using a Dako Envision+ system with dextran polymers conjugated with horseradish peroxidase (Dako), as previously described.^{12 (link)} First, sections were stained with anti-CD68 antibodies for 30 min at room temperature, generating a brown color. Denaturing solution (BioCare Medical-CA, USA) was added for 5 min at room temperature for elution during double staining. Antigen retrieval was performed by heat treatment for 45 min with HIER T-EDTA Buffer (Dako). After incubation, sections were reacted with phospho-STAT1- or c-Maf-specific reagents using dextran polymers conjugated with horseradish peroxidase (Dako) overnight at 4 °C, using a Vina Green Chromogen Kit (BioCare Medical-CA), which produced green staining. Finally, slides were washed in Wash Buffer (Dako) for 3 min. Sections were counterstained with hematoxylin. The antibodies used in this study are listed in Supplementary Table 1.

Shikanai S., Yamada N., Yanagawa N., Sugai M., Osakabe M., Saito H., Maemondo M, & Sugai T. (2023). Prognostic Impact of Tumor-Associated Macrophage-Related Markers in Patients with Adenocarcinoma of the Lung. Annals of Surgical Oncology, 30(12), 7527-7537.

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Histological Analysis of Bone Tissues

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The samples were fixed in 10% neutral buffered formalin and embedded in paraffin wax, which was followed by serial dehydration using ethanol and clearance using xylene. For bone tissues, samples were decalcified with 10% EDTA (pH 7.4) after formalin fixation and delipidation using ethanol. Sections cut into 4-μm thickness were used for Safranin-O/fast green/hematoxylin staining (Saf-O), Alcian blue (pH 1.0) staining (AB), tartrate-resistant acid phosphatase (TRAP) staining, and immunohistochemistry. The information regarding the antibodies and reaction conditions is listed in Additional file 2. After reaction with HPR or AP conjugated antibodies, positive signal color was developed with Histofine Simple stain DAB (NICHIREI BIOSCIENCES, Tokyo, Japan), Vina Green Chromogen Kit (BIOCARE MEDICAL, CA, USA), or ImmPACT Vector Red AP Substrate Kit (VECTOR LABORATORIES, CA, USA). Positively stained areas were measured using ImageJ (National Institutes of Health, MD, USA), ImageScope (Leica Microsystems, Wetzlar, Germany), and BZX-700 (Keyence, Osaka, Japan).

Chijimatsu R., Miwa S., Okamura G., Miyahara J., Tachibana N., Ishikura H., Higuchi J., Maenohara Y., Tsuji S., Sameshima S., Takagi K., Nakazato K., Kawaguchi K., Yamagami R., Inui H., Taketomi S., Tanaka S, & Saito T. (2021). Divergence in chondrogenic potential between in vitro and in vivo of adipose- and synovial-stem cells from mouse and human. Stem Cell Research & Therapy, 12, 405.

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Comprehensive Immunohistochemistry Analysis

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Tissues were paraffin embedded and processed following standard protocols and imaged with a Zeiss CLSM 700 confocal micro-scope. For Lung and Tumor H&E, hematoxylin (VWR, 95057–844) was stained followed by Eosin (VWR, 95057–848). The following antibodies were used for Immunohistochemistry: Rabbit anti-mouse CD8α (D4W2Z) 1:400 (Cell Signaling, 98941), anti-mouse CXCL5 1:200 (LsBio, LS-C104413), rat anti-mouse Ly6g 1:100 (Abcam, ab25377), rabbit anti-mouse β-catenin 1:100 (Cell signaling, 8480). Antigen retrieval was performed by incubation in BioCare’s Nexgen Decloaker chamber for 20 minutes at 90°C. Anti-rabbit polymers were used as secondary antibodies and Vina Green Chromogen Kit (Biocare Medical, BRR 807 AH) was used as substrate for CD8. For CXCL5, LY6G, and β-catenin, anti-rat or rodent polymers were used as secondary antibodies, and Warp Red Chromagen Kit (Biocare Medical, 901-WR806–081017) was used as substrate. ImageJ software was used for quantification.

DeVito N.C., Sturdivant M., Thievanthiran B., Xiao C., Plebanek M.P., Salama A.K., Beasley G.M., Holtzhausen A., Novotny-Diermayr V., Strickler J.H, & Hanks B.A. (2021). Pharmacological Wnt ligand inhibition overcomes key tumor-mediated resistance pathways to anti-PD-1 immunotherapy. Cell reports, 35(5), 109071.

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Comprehensive Immunohistochemistry Analysis

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Dual Immunohistochemistry for Iba1 and PDIA3

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The PT Link (Dako) was used to depara nize and rehydrate the sections and unmask antigen sites. Slides were immersed in 10 mM citrate buffer, pH 6.0, for 10 min at 97°C and then cooled and washed in TBS. Endogenous peroxidase activity was inhibited by incubating the slides with Peroxide Block (ScyTek Laboratories, Utah, USA) for 7 minutes. After washing in distilled water and then in TBS, sections were treated with Avidin-Biotin Kit (Biocare-Medical) to reduce background staining caused by endogenous biotin. After 3 washes in TBS, nonspeci c binding was blocked by 10 minutes incubation with Background Punisher (Biocare-Medical). Sections were incubated over-night at 4°C with Goat Anti-Human Iba1 polyclonal antibody (Novus Biologicals) 1:250, washed extensively in TBS and subsequently incubated with Ultratek HRP kit (ScyTek Laboratories) and the reaction was revealed by incubation with 3,3'-diaminobenzidine (Biocare-Medical). The same slides were washed extensively in distilled water and TBS and blocked with Background Punisher (Biocare-Medical). The rabbit anti-PDIA3 antibody 1:500 was incubated over-night at 4°C and after 3 washes in TBS, slides were incubated with the MACH 2 Rabbit HRP-Polymer (Biocare-Medical). The PDIA3-associated signal was then revealed by incubation with Vina Green chromogen kit (Biocare-Medical).

Chiavari M., Ciotti G.M.P., Canonico F., Altieri F., Navarra P., & Lisi L. (2020). PDIA3 Reduce Glioma-associated Macrophage/microglia Pro-tumor Activation trough IL-6-STAT3-PDIA3 Pathway.

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