Sections of resected tumor tissue (4 μm thick) from the 21 patients were prepared by fixing in formalin and embedding in paraffin. Mouse monoclonal antibodies for Nrf2 (Abcam, ab‐62352), Ki‐67 (Abcam, ab‐21700), p53 (Abcam, ab‐131442), and β‐catenin (Abcam, ab‐16051) were used for staining.23 Staining results were used to separate tumors into a group with low expression of anti‐Nrf2, anti‐Ki‐67, anti‐p53 antibody, and anti‐β‐catenin (<30% of cells with positive staining) and a group with high expression (≥30% of cells with positive staining).23
Ab21700
Manufactured by Abcam
Sourced in United States
Ab21700 is a monoclonal antibody that recognizes the human ERBB2 protein. This antibody is suitable for use in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab205718, by Abcam (3 mentions)
Ab16051, by Abcam (1 mentions)
Ab131442, by Abcam (1 mentions)
Ab62352, by Abcam (1 mentions)
Ab288151, by Abcam (1 mentions)
Ab168809, by Abcam (1 mentions)
Ab32499, by Abcam (1 mentions)
Ab227828, by Abcam (1 mentions)
Ab137550, by Abcam (1 mentions)
Ab6721, by Abcam (1 mentions)
Digital light microscope, by Olympus (1 mentions)
Ab272692, by Abcam (1 mentions)
Ab32572, by Abcam (1 mentions)
Neutral resin, by Solarbio (1 mentions)
Phosphate buffered saline (pbs), by Solarbio (1 mentions)
A microscope, by Olympus (1 mentions)
Hematoxylin, by Solarbio (1 mentions)
Hrp polymer, by Biocare Medical (1 mentions)
St5010 autostainer xl, by Leica camera (1 mentions)
3 3 diaminobenzidine substrate, by Thermo Fisher Scientific (1 mentions)
10 protocols using ab21700
1
Immunohistochemical Profiling of Tumor Markers
2
Immunohistochemical Analysis of ADAR1, Ki-67, Caspase-3, Keap1, and Nrf2
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Tissues were fixed in formalin and embedded in paraffin. Four-μm-thick sections were mounted on poly-l -lysine-coated slides. Afterwards, the slides were dewaxed in xylene and rehydrated with a gradient of ethanol and distilled water. To quench endogenous peroxidase activity, the sections were exposed to 3% hydrogen peroxide for 10 min at room temperature, followed by antigen retrieval. Thereafter, incubation with primary antibodies against ADAR1 (1/100; ab168809; Abcam, USA), Ki-67 (1/100; ab21700; Abcam), Caspase-3 (1/100; ab32499; Abcam), Keap1 (1/200; ab227828; Abcam), and Nrf2 (1/100; ab137550; Abcam) was conducted at 4 °C overnight. Then, the sections were probed with HRP anti-rabbit IgG antibody (1/200; ab288151; Abcam), stained with DAB, and nucleated with hematoxylin, followed by dehydration with a gradient of ethanol and sealing with neutral gum. Protein expression was quantified with ImageJ software.
3
Immunohistochemical Detection of Ki67
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Paraffin sections of tumor tissues were deparaffinized, and incubated with 3% H2O2 (diluted in methanol) at room temperature for 10 min. The antigen was retrieved in citrate buffer, and sections were blocked with 5% BSA, followed by incubation with primary antibody rabbit anti-mouse Ki67 (1: 1, ab21700, Abcam) overnight at 4 °C. Afterwards, the sections were reacted with HRP-conjugated secondary antibody anti-rabbit IgG polyclonal goat antibody (1: 500, ab6721, Abcam) at 37 °C for 1 h. The sections were subjected to color development using DAB, counterstained with hematoxylin, and observed under a microscope [32] (link).
4
Immunohistochemical Analysis of Ki-67 in Tumor Tissues
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The tumor tissues were fixed in 4% paraformaldehyde at room temperature for 24 h, embedded in paraffin and then sliced into 4-µm slices. Next, these slices were subjected to drying, deparaffinization and rehydration. The slices were restored with sodium citrate and incubated with 3% H2O2 for 10 min. Afterward, the slices were blocked in BSA and then subjected to incubation with primary antibody including Ki-67 (Abcam, ab21700, 1:200) overnight at 4°C. On the second day, secondary antibody (Abcam, ab205718, 1:2000) was employed to stain sections at room temperature for 2 h. Subsequently, the sections were stained with DAB mixture (Solarbio, Beijing, China), followed by counterstained with hematoxylin. The tissue sections after staining were examined using Image Pros Plus 5.0 software (Silver Springs, MD, USA).
5
Investigating C-PC and Radiation Therapy for Tumor Growth
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12 BALB/c mice were purchased and injected with CT-26 cells. When the tumors reached 50–70 mm3, the mice were randomly divided into 4 groups (n = 3) including PBS (no-treatment), C-PC, Radiation therapy (RT), and C-PC + RT. The treatment approaches were the same as section 2.6. The mice were sacrificed after 11 days and the tumors were harvested. Immunohistochemistry (IHC) was done according to previous studies45 . Briefly, the tumors were fixed with 10% formalin and then, processed by employing an automatic tissue processor (Sakura, Japan). Then, the paraffin-embedded specimens were processed according to previous studies46 to be stained with anti-Ki-67 antibody (ab21700, Abcam, USA). Immuno-positive cells were quantified at random microscopic fields at ×400 magnification by an expert pathologist. A digital light microscope (Olympus, Tokyo, Japan) was used to capture the photographs.
6
Colonic Immunohistochemistry and Immunofluorescence
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Immunohistochemistry of colonic sections was performed as described previously [23 (link)]. After microwave antigen retrieval, the colonic slides were incubated overnight with primary antibodies of Ki-67 (Abcam, ab21700, Cambridge, UK) and proliferating cell nuclear antigen (PCNA) (Abcam, ab29, Cambridge, UK) at 4 °C. For immunofluorescence staining, colonic slides were incubated overnight with primary antibodies of β-catenin (Abcam, ab32572, Cambridge, UK) and Muc-2 (Abcam, ab272692, Cambridge, UK) at 4 °C.
7
Immunohistochemical Analysis of Xenograft Tumor Tissue
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After collecting xenograft tumor tissues, 4% paraformaldehyde was used to fix the tissues for 2 days, and then, paraffin was used to embed them. The sections were blocked in 1% BSA for 20 min and subsequently cultured with primary antibody against Ki67 (1:20, ab21700, Abcam) at 4 °C overnight. Next, A HSP-labeled Goat Anti-Rabbit IgG H&L (1:500, ab205718, Abcam) was applied to incubate the sections for 30 min at room temperature. Later, hematoxylin was used to counterstain the slides. The images were taken under microscopy (Nikon Microsystems, Tokyo, Japan).
8
Immunohistochemical Analysis of Ki67 in Xenograft Tumors
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Formalin-fixed, paraffin-embedded xenograft tumors were sectioned at 5 μm. Following dewaxing and rehydration, the sections were treated with CPBS buffer (Solarbio) for antigen restoration. After washing with PBS (Solarbio) and blocking with 10% goat serum (Abcam), the sections were probed with anti-Ki67 antibody (ab21700; Abcam), followed by incubation with a secondary antibody (ab205718; Abcam). Then, the sections were counterstained with hematoxylin (Solarbio) after staining with DAB (Leagene). Finally, the sections were observed using a microscope (Olympus) after sealing with neutral resin (Solarbio).
9
Immunohistochemical Analysis of Ki67 in Tissues
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For immunohistochemistry, the tissues (liver, kidney, lung, and tumor) were fixed overnight in 10% formalin, embedded in paraffin and cut into 5 μm sections. The tumor samples were stained with Ki67 (ab21700, Abcam). After overnight incubation, the slides were washed and incubated with the secondary antibody (HRP-Polymer, Biocare Medical) for 30 min at room temperature. The slides were washed three times and stained with 3, 3’-diaminobenzidine (DAB) substrate (Thermo Fisher Scientific). The slides then were counterstained with hematoxylin and mounted with a mounting medium. In addition, the hematoxylin–eosin (HE) staining was performed on paraffin-embedded tissue sections by the ST5010 Autostainer XL (Leica). Images were obtained by the Aperio Image Scope Viewer (Leica). Quantification by counting of Ki67+ cells number in random area per sample.
10
Xenograft Tumor Characterization by IHC
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The xenograft tumors were fixed with 4% neutral-buffered formalin, dehydrated, embedded in paraffin, and sectioned at 5 µm intervals. Proliferation and invasion of the xenograft tumors were evaluated by immunohistochemistry using anti-Ki-67 (ab21700; Abcam; dilution 1:1) and anti-MMP9 (ab228403; Abcam; dilution 1:5,000) antibodies, respectively. Briefly, paraffin-embedded sections were dewaxed, heated in 0.01 M of sodium citrate buffer, and treated with methanolic H2O2, followed by the incubation with anti-Ki-67 and anti-MMP9. Immunoreactive signals were detected using 3,3′-diaminobenzidine substrate (Thermo Fisher Scientific), and then the sections were counterstained with hematoxylin and mounted. Apoptosis of the tumor cells was assessed by staining paraffin-embedded sections with the TUNEL Assay Kit (Cat. ab206386; Abcam) as per the accompanying guidance.
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