Anti-human Ki-67 antibody (DAKO) and anti-human ABCB1 antibody (Cell Signaling #13342).
Anti human ki 67 antibody
Manufactured by Agilent Technologies
Sourced in United States
The Anti-human Ki-67 antibody is a laboratory reagent used to detect the presence of the Ki-67 protein, which is a marker of cellular proliferation. This antibody can be used in various immunohistochemical and immunocytochemical techniques to identify and quantify proliferating cells.
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Lab products found in correlation
Inform software, by PerkinElmer (1 mentions)
Vectra polaris automated quantitative pathology imaging system, by PerkinElmer (1 mentions)
Refine detection kit, by Leica (1 mentions)
Bond max, by Leica (1 mentions)
E cadherin antibody, by Santa Cruz Biotechnology (1 mentions)
Target retrieval solution ph 9, by Agilent Technologies (1 mentions)
Herceptest 2, by Agilent Technologies (1 mentions)
Envision system, by Agilent Technologies (1 mentions)
Clone mib 1, by Agilent Technologies (1 mentions)
Hpa012323, by Merck Group (1 mentions)
Anti mouse cd45 antibody, by Abcam (1 mentions)
Accutase, by STEMCELL (1 mentions)
Tunel staining, by Roche (1 mentions)
Alexa fluor 488 goat anti rabbit igg, by Abcam (1 mentions)
Alexa fluor 594 goat anti mouse igg, by Abcam (1 mentions)
8 protocols using anti human ki 67 antibody
1
Evaluating Proliferation and ABC Transporter
2
Quantifying Ki67 Expression in Breast Cancer
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Formalin-fixed, paraffin-embedded biopsy specimens and parallel surgical excisions from patients with early stage breast cancer were subjected to immunohistochemistry (IHC) staining using anti-human Ki67 antibody (Dako, clone 30-9). Results were analyzed using the Vectra® Polaris™ Automated Quantitative Pathology Imaging System and InForm software (PerkinElmer) (1 × 1 field, 20× resolution).
3
Ki-67 Immunohistochemistry Tumor Assessment
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Whole section Ki-67 expression in tumor specimen was assessed by immunohistochemistry using the Leica Bond Max (Leica Biosystems, Wetzlar, Germany) with heat-induced epitope regaining. Staining was carried out following the protocol indicated by the International Ki-67 in Breast Cancer Working Group [25 (link)]. Slides were stained at room temperature for 1 h with the primary antibody (monoclonal mouse, anti-human Ki-67 antibody; DAKO, Carpinteria, CA, USA) at a dilution of 1:100. Then, the primary antibody was detected using the Refine Detection Kit (Leica Biosystems, Wetzlar, Germany), which contains a rabbit anti-mouse IgG secondary antibody and anti-rabbit poly-HRP IgG antibody, and 3,3′-diaminobenzidine was used as a chromogen. The slides were finally counterstained with hematoxylin. The Ki-67 proliferation index was the percentage of positively stained nuclei scored by manual scoring, which calculated the percentage of positively stained nuclei within three high-powered fields (×40 magnification) randomly identified throughout the tumor, assuring no less than 1000 nuclei were counted [26 (link)].
4
Immunohistochemical Evaluation of MET, HGF/SF, and Proliferation Markers
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To determine MET expression status by immunohistochemistry (IHC), monoclonal antibody (mAb) for MET, MET4 mAb,26 was used. Epitope sequence of MET4 revealed that DVLPEFR on amino acid residue 236–242 was the specific recognition site of MET. To retrieve MET antigen, tissue slide sections were treated at 98°C for 20 min in Target Retrieval Solution, pH9 (DAKO, Santa Clara, CA). ENVISION+ System (DAKO) was used for the secondary antibody reaction and DAB+ was used for color development. In the final step, the nuclei were lightly counterstained with Meyer hematoxylin.
Regarding the tissue staining of HGF/SF, anti‐HGF/SF mAb (clone 7‐2) was purchased from Enzo Life Sciences (Ann Arbor, MI, USA) and the samples were stained according to the company's instruction. Antigen retrieval was performed by boiling the sample slides in the 10 mM citrate buffer (pH 6) in a microwave for 7–10 min.
For the evaluation of Her2/neu IHC status HercepTest II (DAKO) was used. For the evaluation of tumor cell proliferation, we performed Ki‐67 staining using anti‐human Ki‐67 antibody (M7240, DAKO). E‐cadherin expression in the proliferative margin of tumor cells was also investigated using E‐cadherin antibody (H‐108, Santa Cruz Biotechnology, CA, USA).
Regarding the tissue staining of HGF/SF, anti‐HGF/SF mAb (clone 7‐2) was purchased from Enzo Life Sciences (Ann Arbor, MI, USA) and the samples were stained according to the company's instruction. Antigen retrieval was performed by boiling the sample slides in the 10 mM citrate buffer (pH 6) in a microwave for 7–10 min.
For the evaluation of Her2/neu IHC status HercepTest II (DAKO) was used. For the evaluation of tumor cell proliferation, we performed Ki‐67 staining using anti‐human Ki‐67 antibody (M7240, DAKO). E‐cadherin expression in the proliferative margin of tumor cells was also investigated using E‐cadherin antibody (H‐108, Santa Cruz Biotechnology, CA, USA).
5
Tumor Cell Proliferation and Immunophenotyping
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Freshly excised tumors were manually minced before enzymatic disaggregated for 2 hrs with Accutase (Stem Cell technologies, Vancouver, CA) and pooled. Cell proliferation was assessed by quantification with Ki-67 immunohistochemistry on cytospin sections of the disaggregated tumors with a anti-human Ki-67 antibody (1:300, clone MIB-1) (Dako, CA, USA). Positive cells were scored by visual examination of 10 randomly chosen fields containing at least 100 cells. For FACS analysis of the tumor-derived cells, anti-human CSF-1R antibody (1:200, HPA012323) (SIGMA-Aldrich, Milan, Italy) and anti-mouse CD45 antibody (1:100, ABCAM, Cambridge, UK) were used
6
Immunohistochemical Analysis of Tumor Samples
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Sections (2-μm thick) from FFPE human tumor xenografts and lymph node biopsies obtained from consenting patients at the time of diagnostic work-up were stained with hematoxylin and eosin (H&E) or with anti-human Ki-67 antibody (Dako, Milan, Italy, EU), -pAKT (S473), -pAKT (T308), -pERK1/2 or -pS6 (Cell Signaling) antibodies. Chemosensitive and chemorefractory patients were defined according to the criteria of German Hodgkin Lymphoma Study Group59 (link). Study was approved by the by the institutional Ethical Committee of the Humanitas Clinical and Research Center. Tumor necrosis was detected via TUNEL staining (Roche, Milan, Italy, EU). Cryostat sections (4-μm thick) of in vivo biotinylated tumor nodules were processed as previously described28 (link). The sections were examined under a light microscope (IX51; Olympus, Tokyo, Japan). Image analysis was performed using ImageJ software60 (link).
7
Immunohistochemical analysis of Ki-67
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Immunohistochemical staining was conducted on 4‐μm‐thick sections. Tumour sections were subjected to deparaffinize and rehydrate with gradient ethanol solutions. After being immersed in antigen retrieval solution and heated, sections were incubated with anti‐human Ki‐67 antibody (Dako) for 1 hour and counterstained with haematoxylin (BASO, China) for 2 minutes.
8
Immunohistochemistry of Sheep Lung Tissue
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Immunohistochemistry was performed on frozen tissue of sheep lung tissue. Tissue sections were stained using previously described methods. Primary antibodies used were: anti-human KCNN4/K Ca 3.1 (rabbit polyclonal, 1:200; Lifespan Biosciences, Seattle, WA), anti-a-smooth muscle actin (a-SMA; mouse monoclonal, 1:800; Sigma-Aldrich), anti-human Ki-67 antibody (mouse polyclonal, 1:100; DAKO, Santa Clara, CA). For fluorescent staining, secondaries used were Alexa Fluor 488 goat anti-rabbit IgG and Alexa Fluor 594 goat anti-mouse IgG (both Abcam, Cambridge, UK). For peroxidase staining, goat antirabbit or mouse IgG (both Abcam) were used.
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