Tissue samples were fixed in 4% buffered formaldehyde, dehydrated, embedded in paraffin, and sectioned at 2 µm. H&E staining was performed according to standard laboratory protocols. Immunohistochemical stainings were performed on a Ventana BenchMark XT system (Roche Diagnostics). The following primary antibodies were used: SALL4 (ab57577, abcam, 1:50), Ki67 (SP6, Cell Marque, 1:750), SOX2 (ab97959, abcam, 1:1000), MAP2C (M4403, Sigma–Aldrich, 1:3000), CD56 (MSK006, Zytomed, 1:2000), Synaptophysin (M7315, Dako, 1:500). As a chromogen, 3,3ʹ-diaminobenzine (DAB) was used.
Ki67 sp6
Manufactured by Cell Marque
Sourced in United States
Ki67 (SP6) is a rabbit monoclonal antibody used for the immunohistochemical detection of the Ki67 protein, which is a cellular marker for proliferation. Ki67 is expressed during all active phases of the cell cycle (G1, S, G2, and mitosis), but is absent in resting cells (G0).
Automatically generated - may contain errors
Lab products found in correlation
Ventana benchmark xt system, by Roche (1 mentions)
M4403, by Merck Group (1 mentions)
Ab57577, by Abcam (1 mentions)
Ab97959, by Abcam (1 mentions)
M7315, by Agilent Technologies (1 mentions)
Starr trek universal hrp detection system, by Biocare Medical (1 mentions)
Labvision autostainer 480s, by Thermo Fisher Scientific (1 mentions)
Cyp1b1, by Abcam (1 mentions)
Cd44 clone 156 3 c11, by Cell Signaling Technology (1 mentions)
4 protocols using ki67 sp6
1
Tissue Immunohistochemistry Protocol
2
Immunohistochemical Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical (IHC) techniques were performed according to Agustina et al., [8 ]. IHC staining on the samples was performed manually using a labelled streptavidin-biotin immunoperoxide complex method, using the Starr Trek Universal HRP Detection system (Biocare Medical, Concord, CA, USA). Each cell block was cut into 4-µm slices and examined on L-lysine coated glass slides and baked at 60°C for one hour on a standard histology hotplate. Deparaffinized using xylene and rehydrated using an alcohol solution than brought to water. Antigen retrieval used a decloaking chamber (DC2008INTL, Biocare Medical, USA) in EDTA (pH 8.0), followed by cooling at room temperature for 20 minutes. Sections were then treated to block endogenous peroxidase, stained with primary antibodies, and incubated for 1 hour at room temperature. Detection was done by horseradish peroxidase polymer-based detection system (Biocare Medical) and diaminobenzidine chromogen and counterstained with haematoxylin. The primary antibodies were E-cadherin (G10) sc-8426 from Santa Cruz Biotechnology, inc (Santacruz, CA) and KI-67(SP6) from Cell Marque (Rocklin, CA, USA).
3
Immunohistochemical Staining of Tissue Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue samples were incubated in 4% formalin overnight and subsequently embedded in paraffin. For tissue analysis, 3–5 μm sections were cut, de-paraffinized, and antigen retrieval was performed using either citrate at pH 6.0, or ethylenediaminetetraacetic acid (EDTA) at pH 9.0 for 20 min. Washing steps were performed using phosphate-buffered saline. Primary antibodies were purchased from Cell Signaling (Cleaved Caspase-3, #Asp175, dilution 1:100) and Cell Marque (Ki67, #SP6, dilution 1:100). Corresponding secondary antibody detection kits were used from Histofine® Simple stain and stained on an automated stainer (LabVision Autostainer 480S; Thermo Fisher).
4
Immunohistochemical Protein Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein staining using IHC was assessed using 4-μm-thick paraffin-embedded tissue sections, as described previously [28] (link), using antibodies against AHR (dilution 1:500) (Santa Cruz, Texas, USA), CYP1B1 (dilution 1:500) (Abcam, MA, USA), CD44 (clone 156-3-c11) (dilution 1:800) (Cell Signaling Technology, MA, USA) and Ki-67 (SP6) (Cell Marque Corporation, CA, USA). IHC staining was carried out by adding 100 µL of the chromogen DAB + diluted 1:50 in substrate buffer [EnVision + Dual Link System-HRP (DAB + )] for 10 min. Finally, tissue specimens were washed in phosphate-buffered saline (PBS), the nuclei were counterstained with haematoxylin, and the specimens were mounted using Permount® for examination by light microscopy. The stained area fractions were evaluated using ImageJ software (National Institutes of Health, MD, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!