Formalin-fixed, paraffin-embedded sections of surgical specimens from patients with ESCC were deparaffinized and rehydrated. Antigen retrieval was performed in 10 mM sodium citrate buffer (pH 6.0) using an autoclave for 5 min. Sections were stained with hematoxylin and eosin (H/E) or anti-PITX1 antibody (HPA008743, Sigma-Aldrich, Inc.). A diaminobenzidine staining procedure was performed using the ImmPACTTM DAB peroxidase substrate kit (Vector Laboratories, Burlingame, CA), and hematoxylin was used for counterstaining. Ki67 was detected using anti-Ki67 antibody (MIB-1, Dako, Glostrup, Denmark) after antigen retrieval.
Anti ki67 antibody mib 1
Manufactured by Agilent Technologies
Sourced in Denmark
The Anti-Ki67 antibody (MIB-1) is an immunohistochemical reagent used to detect the Ki67 protein, which is a cellular marker of proliferation. The antibody is designed for use in paraffin-embedded tissue sections and can be used to identify and quantify the proportion of actively proliferating cells in a tissue sample.
Automatically generated - may contain errors
Lab products found in correlation
Immpact dab peroxidase substrate kit, by Vector Laboratories (1 mentions)
Anti cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Glyceraldehyde 3 phosphate dehydrogenase 2118, by Cell Signaling Technology (1 mentions)
Sc 79, by Santa Cruz Biotechnology (1 mentions)
Anti bcl 2 antibody, by Santa Cruz Biotechnology (1 mentions)
Histostain plus kit, by Zymo Research (1 mentions)
4 protocols using anti ki67 antibody mib 1
1
PITX1 and Ki67 Expression in ESCC
2
Immunohistochemical Staining of BCBM
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IHC staining was performed as described previously (9). Anti-Ki67 antibody (MIB-1) was from DAKO. Anti-Cleaved Caspase 3 antibody was from Cell Signaling Technology. Anti-HER2 (OP15) antibody was purchased from MilliporeSgma. Quantification of Ki67 and Cleaved Caspase 3 was conducted using the Image J software with ImmunoRatio plugin. Sections of BCBM tissue from 2 animals per condition were imaged and 4–6 randomly chosen regions of interest per section were used for quantification.
3
Western Blotting Antibody Panel
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The following primary antibodies were used for western blotting. Anti–cleaved caspase 3 (CASP3; #9664), cleaved CASP8 (#9496), cleaved poly‐ADP‐ribose polymerase (PARP) (#9546), TFEB (#4240), lamin (#2032), and glyceraldehyde‐3‐phosphate dehydrogenase (#2118) were purchased from Cell Signaling Technology. GAA (ab137068), lysosomal associated membrane protein 2 (LAMP2; ab25631), and sequestosome 1 (SQSTM1)/p62 (ab56416) were purchased from Abcam. Light chain 3 (LC3; NB110‐57180) was purchased from Novus Biologicals. Anti–Ki‐67 antibody (MIB‐1) was purchased from Dako.
4
Immunohistochemical Analysis of Angiogenic Factors
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Additional sections placed on 3-aminopropyltriethoxysilane (Sigma, USA)-coated slides were stained using the streptavidin-biotin-peroxidase complex technique (Histostain Plus Kit; ZYMED, USA) with monoclonal anti-VEGF antibody (SC-7269; Santa Cruz Biotechnology, USA), anti-Bcl-2 antibody (SC-7382; Santa Cruz Biotechnology), anti-Ki-67 antibody (MIB-1; Dako, Denmark), and polyclonal anti-FGF antibody (SC-79; Santa Cruz Biotechnology). Antigen retrieval was performed by heating the sections in citrate buffer (pH 6.0) for 20 min in a microwave oven at 600 W. Aminoethylcarbazole or 3,3-diaminobenzidine in H2O2 were used as chromogens with incubation for 10 min (controlled by visual observation under a microscope). The sections were counterstained with Mayer's hematoxylin for 1 min, rinsed with tap water, and mounted with an aqueous mounting medium. The percentages of the total area of the positive cells were determined using a microscopy image analysis system (Bs200P Image Analysis System; BAB Software, Turkey). A total of 10 high-power fields were randomly chosen and analyzed at 400× magnification.
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