RCC tissue microarrays with 180 samples were purchased from Outdo Biotech Co., Ltd (Shanghai, China). The primary antibodies against PUF60, TERT, Ki67, cyclin D1 and PCNA were diluted 1:100, and then incubated at 4°C overnight in a humidified container. After three times washes with PBS, the tissue slides were treated with a non-biotin horseradish peroxidase detection system according to manufacturer's instructions (Dako). IHC scores were calculated by image pro plus 6 software according to the manufacturer's instructions.
Non biotin horseradish peroxidase detection system
Manufactured by Agilent Technologies
Sourced in Denmark
The Non-biotin horseradish peroxidase detection system is a laboratory equipment product that utilizes horseradish peroxidase as the detection enzyme, without the use of biotin. It is designed to provide a sensitive and reliable method for detecting target analytes in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Sc 130867, by Santa Cruz Biotechnology (1 mentions)
Abd29, by Santa Cruz Biotechnology (1 mentions)
Hbre duc140sur 01, by Shanghai Outdo Biotech Co. (1 mentions)
Dab substrate, by Agilent Technologies (1 mentions)
Rabbit polyclonal anti adamts5, by Abcam (1 mentions)
Mouse monoclonal anti igfbp5, by Abcam (1 mentions)
Ab230513, by Abcam (1 mentions)
Ab16066, by Abcam (1 mentions)
Ab28364, by Abcam (1 mentions)
19 protocols using non biotin horseradish peroxidase detection system
1
Immunohistochemical Analysis of RCC Tissues
2
Immunohistochemical Analysis of Osteosarcoma
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This procedure was described previously [30 (link), 35 (link), 37 (link)]. Immunohistochemical analysis was performed on prepared3-μm sections. The primary antibodies against BRD7, Cdh1 and Cdc20 were diluted 1:250, 1:100 and 1:500, respectively, and were incubated at 4°C overnight in a humidified container. After washing with PBS three times, the tissue slides were treated with a non-biotin horseradish peroxidase detection system according to the manufacturer's instructions (Dako). IHC staining was evaluated by two independent pathologists who are experts in osteosarcoma pathology (Dr. An-Jia Han and Dr. Hui-Juan Shi). The protein expression levels of BRD7, Cdh1 and Cdc20 were classified as high level when the tumor tissue had more than 10% of cells positive for staining; otherwise, the expression levels were classified as low.
3
Immunohistochemical Analysis of pAKT and PIK3CA in Gastric Carcinoma
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Formalin-fixed, paraffin-embedded tissue specimens from gastric carcinoma patients were cut in 5 μm sections. Briefly, after deparaffinized in xylene and rehydrated, the tissue slides were then treated with 3% hydrogen peroxide, and the antigens were retrieved in sodium citrate buffer (pH 6.0) using a microwave oven. After 1 hour of preincubation in goat serum to prevent nonspecific staining, the specimens were incubated with primary antibodies overnight at 4°C. The tissue slides were treated with a non-biotin horseradish peroxidase detection system according to the manufacturer’s instructions (DAKO, Glostrup, Denmark). Two different pathologists evaluated the results of IHC. Both the extent and intensity of immunostaining were taken into consideration. The intensity of staining was scored from 0 to 3, and the extent of staining was scored from 0% to 100%. The final score of each staining was obtained by multiplying the two scores. The pAKT expression was recognized as higher level, if the score was higher than 1.5; if the score was 1.5 or less, the case was classified as low expression. PIK3CA expression was considered high if the score reached 0.5.
4
Immunohistochemical Analysis of RPA3 and BRCC3 in FFPE Tissue
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Formalin‐fixed, paraffin‐embedded tissue specimens were analysed by immunohistochemistry according to previous reports 25 , 26 . Briefly, rabbit polyclonal antibodies against RPA3 (working dilution 1:100; PA5‐21277; Pierce) and BRCC3 (working dilution 1:200; PA5‐20426; Pierce) were used for immunohistochemistry (IHC) assays, and a non‐biotin horseradish peroxidase detection system (DAKO, Glostrup, Denmark) was used to detect the expression level of the protein of interest. Both the extent and intensity of immunostaining were taken into consideration when analysing the data. The intensity of staining was scored from 0 to 3, and the extent of staining was scored from 0% to 100%. The final quantitation of each stain was obtained by multiplying the two scores. RPA3 expression was classified as high expression if the score was higher than 1.5, whereas scores of 1.5 or less indicated low expression.
5
Immunohistochemical Analysis of Tongue Cancer
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Human tongue cancer specimens were cut into 4-μm sections. The sections were dried at 62°C for 2 h and then deparaffinized in xylene and rehydrated using a series of graded alcohol washes. The tissue slides were then treated with 3% hydrogen peroxide in methanol for 15 min to quench endogenous peroxidase activity and antigen retrieval then performed by incubation in 0.01 M sodium cirate buffer (pH 6.0) and heating using a microwave oven. After a 1 h preincubation in 10% goat serum, the specimens were incubated with primary antibody overnight at 4°C. The tissue slides were treated with a non-biotin horseradish peroxidase detection system according to the manufacturer's instruction (DAKO, Glostrup, Denmark). Two different pathologists evaluated the immunohistological samples. The intensity of immunostaining was taken into consideration when analyzing the data. The intensity of staining was scored from 0 to 3 and the expression was classified as high if the score was ≥2, and as low if the score was ≤1.
6
Immunohistochemical Analysis of PUF60 in Bladder Cancer
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Bladder cancer tissue microarrays with 56 samples were purchased from Outdo Biotech Co., Ltd. (Shanghai, China). Bladder cancer tissue microarrays with 13 pairs of samples were purchased from Alenabio Biotechnology Co., Ltd. (Xi’an, China). The primary antibodies against PUF60 were diluted 1:100, and then incubated at 4°C overnight in a humidified container. After three washes with PBS, the tissue slides were treated with a non-biotin horseradish peroxidase detection system according to manufacturer’s instructions (Dako). IHC scores were evaluated. IHC H-score = Σ(PI × I) = (percentage of cells of weak intensity × 1) + (percentage of cells of moderate intensity × 2) + (percentage of cells of strong intensity × 3).
7
Immunohistochemical Analysis of β-Catenin, Serglycin, and P-ERK
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IHC analysis was performed on three sections. Primary antibodies against β-catenin, serglycin and P-ERK were diluted 1 : 100 and were incubated at 4 °C overnight in a humidified container. After washing with PBS three times, tissue slides were treated with a non-biotin horseradish peroxidase detection system according to the manufacturer's instructions (Dako, Glostrup, Denmark).
8
Immunohistochemical Analysis of SUN2 in Lung Cancer
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This procedure was described previously39 (link)40 (link). The immunohistochemical analysis was performed on 3-μm sections. Primary antibody against SUN2 was diluted 1:2000 and was incubated with the samples at 4 °C overnight in a humidified container. After washing with PBS three times, the tissue slides were treated with a non-biotin horseradish peroxidase detection system according to the manufacturer’s instructions (Dako). The resultant IHC staining was evaluated by two independent pathologists majoring in lung cancer. The protein expression of SUN2 was evaluated using the semiquantitative IRS (immunoreactive score) scale according to Remmele and Stegner41 (link).
9
Immunohistochemical Analysis of Protein Expression
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The sections were dried at 55°C for 2 h and then deparaffinized in xylene and rehydrated using a series of graded alcohol washes. The tissue slides were then treated with 3% hydrogen peroxide in methanol for 15 min to quench endogenous peroxidase activity and antigen retrieval then performed by incubation in 0.01 M sodium cirate buffer (pH 6.0) and heating using a microwave oven. After a 1 h preincubation in 10% goat serum, the specimens were incubated with primary antibody overnight at 4°C. The tissue slides were treated with a non-biotin horseradish peroxidase detection system according to the manufacturer's instruction (DAKO, Glostrup, Denmark). Two different pathologists evaluated the immunohistological samples. The intensity of immunostaining was taken into consideration when analyzing the data. The intensity of staining was scored from 0 to 3 and the expression was classified as high if the score was ≥ 2, and as low if the score was ≤ 1.
10
Immunohistochemical Analysis of Tongue Cancer
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A tissue array containing 84 human tongue cancer specimens was cut into 4-μm sections. The sections were dried at 62°C for 2 h and then deparaffinized in xylene and rehydrated using a series of graded alcohol washes. The tissue slides were then treated with 3% hydrogen peroxide in methanol for 15 min to quench endogenous peroxidase activity and antigen retrieval then performed by incubation in 0.01 M sodium cirate buffer (pH 6.0) and heating using a microwave oven. After a 1 h preincubation in 10% goat serum, the specimens were incubated with primary antibody overnight at 4°C. The tissue slides were treated with a non-biotin horseradish peroxidase detection system according to the manufacturer's instruction (DAKO, Glostrup, Denmark). Two different pathologists evaluated the immunohistological samples.
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