Two step ihc kit

Manufactured by ZSGB-BIO

The Two-Step IHC Kit is a laboratory equipment product designed for immunohistochemistry (IHC) staining procedures. It consists of a primary antibody and a secondary antibody-enzyme conjugate, which work together to detect and visualize specific target proteins in tissue samples.

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Lab products found in correlation

Ab227958, by Abcam (1 mentions) Strataquest software, by TissueGnostics (1 mentions) Ab30682, by Abcam (1 mentions) Ab45174, by Abcam (1 mentions) Ab40793, by Abcam (1 mentions) Df6106, by Affinity Biosciences (1 mentions) Af6283, by Affinity Biosciences (1 mentions) Bs 2130r, by Bioss Antibodies (1 mentions)

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IHC kit (20 citations)

2 protocols using two step ihc kit

Quantitative IHC Analysis of NEK2 in ccRCC

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Immunohistochemical (IHC) analysis was conducted using tissue microarray (TMA) of ccRCC and adjacent normal tissue specimens according to the protocol of the Two-Step IHC Kit (ZSGB-BIO, Beijing, China) using an anti-NEK2 antibody (ab227958, 1:200; Abcam, Cambridge, UK). The specimens of TMA were reviewed by one uropathologist and the most representative areas of renal tumor cells and adjacent tumor stroma were selected for the donor block. The TMAs were made by using a tissue-arraying instrument (Manual Tissue Microarrayer Quick-Ray ®). Cores with 2 mm diameter from per-donor block were diverted into a recipient block microarray.
The StrataQuest software (TissueGnostics, Vienna, Austria) was used to scan and analyze the images to quantify the tumor tissues, para-carcinoma tissues and NEK2 positive cells in the TMA. We multiplied the quantitative value of positive staining intensity (negative = 1, weak = 2, moderate = 3, strong = 4) by percentages of positive staining (0% = 0, ≤25% = 1, 26%–50% = 2, 51%–75% = 3, ≥76% = 4), and the data obtained was weighted and set into four levels (Ⅰ, Ⅱ, Ⅲ, Ⅳ). All results were verified by three experienced independent investigators.

Wang C., Huang Y., Ma X., Wang B, & Zhang X. (2021). Overexpression of NEK2 is correlated with poor prognosis in human clear cell renal cell carcinoma. International Journal of Immunopathology and Pharmacology, 35, 20587384211065893.

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Comprehensive Immunohistochemistry Protocol

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IHC was conducted according to the protocol of the Two-step IHC Kit (ZSGB-BIO). Briefly, 4 µm thick slides of paraffin-embedded tissue specimens were processed for antigen retrieval with citric acid buffer (0.01 M, pH 6.0), and endogenous peroxidase activity was quenched with 3% hydrogen peroxide solution. Then, the slides were blocked with 10% bovine serum albumin and incubated overnight at 4 °C with the following primary antibodies: anti-MED15 (1:50; 11566-1-AP; Proteintech), anti-SREBP1 (1:100; AF6283; Affinity), anti-SREBP2 (1:100; ab30682; Abcam), anti-FASN (1:50; DF6106; Affinity), anti-ACC1 (1:100; ab45174; Abcam), anti-ACLY (1:100; ab40793; Abcam), anti-SCD1 (1:100; #2794; CST), anti-PLK1 (1:50; DF7004; Affinity), and anti-Ki67 (1:200, BS-2130R; Bioss). The slides were incubated with biotinylated goat anti-rabbit IgG (1:200) for 1 h at room temperature. DAB (ZLI-0918, ZSBio, China) was used to visualize the immune complexes, and the sections were counterstained with hematoxylin. Immunostaining intensity was measured using ImageJ software (National Institutes of Health, Bethesda, MD) as previously described [56 (link)].

Hua X., Ge S., Zhang L., Jiang Q., Chen J., Xiao H, & Liang C. (2024). MED15 is upregulated by HIF-2α and promotes proliferation and metastasis in clear cell renal cell carcinoma via activation of SREBP-dependent fatty acid synthesis. Cell Death Discovery, 10, 188.

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