The microarray was incubated with primary antibodies against AKR1C1. Sections were subsequently incubated with the Cell and Tissue Staining Kit HRP-DAB system (R&D Systems, Inc., Minneapolis, MN, USA) according to the manufacturer’s instructions. Formalin-fixed paraffin-embedded H446 tumors were cut with a microtome into 6 μm sections. Antigen retrieval was performed in 10 mM sodium citrate buffer at pH 6.0 for 16 minutes at 96°C–98°C. Slides were incubated with primary antibodies against AKR1C1, P-ERK, and EGFR. Sections were subsequently incubated with the Cell and Tissue Staining Kit HRP-DAB system (R&D Systems, Inc.), according to the manufacturer’s instructions. The positive expression of AKR1C1 was mainly located in the cytoplasm and stained brown. The staining intensity was scored as follows: 0= no color; 1= light yellow; 2= yellow; 3= brown; and 4= dark brown. Positive cells accounted for a percentage score standard: 0= no positive cells; 1=1%–25% positive cells; 2=26%–50% positive cells; 3=51%–75% positive cells; and 4= more than 75% positive cells. A total score of the two lower than 4 was defined as low expression, and a total score of the two higher than 4 was defined as high expression.
Cell and tissue staining kit hrp dab system
Manufactured by R&D Systems
Sourced in United States
The Cell and Tissue Staining Kit HRP-DAB system is a laboratory equipment product designed for the visualization of target proteins, antigens, or other biomolecules in cell and tissue samples. The kit utilizes horseradish peroxidase (HRP) and 3,3'-diaminobenzidine (DAB) as the detection system.
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Lab products found in correlation
7 protocols using cell and tissue staining kit hrp dab system
1
Immunohistochemical Analysis of AKR1C1
2
Chordoma Tissue Immunohistochemistry
3
Immunohistochemical Staining and Microscopy
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PAS and elastica Masson trichrome staining were performed as described above. Immunohistochemical staining was performed with the HRP-DAB System Cell and Tissue Staining Kit (R&D Systems, Minneapolis, MN, USA) and the above-mentioned primary Abs after antigen retrieval in Target Retrieval Solution. Toluidine blue staining and transmission electron microscopy were performed as described above with minor differences in reagents and devices. Information on Abs used is provided in Supplemental Table 2 .
4
Immunohistochemical analysis of U87 tumors
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Formalin-fixed paraffin-embedded U87 tumors were cut with a microtome into 6-μm sections. Antigen retrieval was carried out in 10 mM sodium citrate buffer of pH 6.0 for 16 min at 96–98°C. Slides were incubated with primary antibodies against MMP9, Ki-67 (Boster Bioengineering, Wuhan, China) and NF-κB1. Sections were subsequently incubated with the Cell and Tissue Staining Kit HRP-DAB system (R&D Systems, Minneapolis, MN, USA), according to the manufacturer's instructions. Immunostainings were run with known positive and negative tumor controls, and were blindly evaluated by a pathologist.
5
Immunohistochemical Analysis of Ki-67
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U87 and U251 cells were incubated with the primary antibody anti-Ki-67 (CST). Sections seeded with cells were incubated according to manufacturer’s instructions offered by the Cell and Tissue Staining Kit HRP-DAB system (R&D Systems, Minneapolis, MN, USA).
6
Immunohistochemical Analysis of NFKBIA and NF-κB1
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The sections were deparaffinized in xylene and rehydrated in graded alcohols. Following deparaffinization, antigen retrieval was performed by immersing the sections in 10 mmol citrate buffer (pH 6.0) and heating them twice in a microwave oven (95°C) for 5 min. The sections were incubated with primary antibodies against NFKBIA (Clone ID: EP697; Epitomics; dilution 1:100 overnight at 4°C) and NF-κB1 (Abcam, Tokyo, Japan). The sections were subsequently incubated using the Cell and Tissue Staining kit HRP-DAB system (R&D Systems), according to the manufacturer’s instructions. Immunostainings were performed with known positive and negative tumor controls, and were blindly evaluated by a pathologist.
7
Immunohistochemical Analysis of MMP-2 and MMP-9
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Formalin-fixed paraffin-embedded U-373 tumors were cut with a microtome into 5-μm-thick paraffin sections. Antigen retrieval was carried out in heated 10 mM citrate buffer of pH 6.0 for 10 min at 96°–98°C. Slides were incubated with primary antibodies against matrix metalloproteinase-2 (MMP-2) and MMP-9 (Boster Bioengineering, Wuhan, P.R. China). Corresponding mouse HRP-conjugated secondary antibody was added for 1 h at room temperature. Cells were counterstained with 10 mg/ml DAPI. Sections were subsequently incubated with the Cell and Tissue Staining Kit HRP-DAB system (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.
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