Liver tissues were fixed in 10% neutral buffered formalin, paraffin embedded and cut into 4 μm sections. Sections were H&E stained and assessed for tumour grade by pathologist Professor Robert Goldin (Imperial College London). Necrosis was determined by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) staining (ApopTag Plus Peroxidase In Situ Apoptosis Detection Kit, Milipore). Hepatocyte proliferation was determined by staining with polyclonal rabbit anti-MCM4 antibody at a 1/100 dilution (Abcam, ab84153). PD-1 expressing lymphocytes were identified using a rabbit polyclonal anti PD-1 antibody at 2.5 μg ml−1 (Biorad ahp1706). RAE-1 expression was analysed using a polyclonal goat anti-mouse RAE-1 antibody at 10 μg ml−1 (R&D Systems). HCC markers were stained using anti-Hsp70 at 1 μg ml−1 (EPR16892) and rabbit polyclonals raised against Glypican-3 at 5 μg ml−1 (ab66596) and Anti-Glutamine Synthetase at 1 μg ml−1 (ab73593) from abcam. Slide images were captured using a Nanozoomer slide scanner (Hamamatsu) and analysed using Image J software.
Epr16892
Manufactured by Abcam
Sourced in United States
EPR16892 is a recombinant monoclonal antibody that recognizes the protein Abcam. This antibody is suitable for use in various laboratory applications, including Western blotting and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Ab66596, by Abcam (1 mentions)
Ab73593, by Abcam (1 mentions)
Apoptag plus peroxidase in situ apoptosis detection kit, by Merck Group (1 mentions)
Nanozoomer slide scanner, by Hamamatsu Photonics (1 mentions)
Ab125011, by Abcam (1 mentions)
Ice cold lysis buffer, by Roche (1 mentions)
Epr21151, by Abcam (1 mentions)
Epr16769, by Abcam (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit igg, by Boster Bio (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
2 protocols using epr16892
1
Comprehensive Liver Tumor Analysis
2
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Protein homogenates from A549 cells and H2170 cells were extracted as previously described. Brie y, the cells were lysed for 20 min on ice in ice-cold lysis buffer (Roche). The lysates were centrifuged at 12,000 × g for 20 min at 4•C to obtain a clear lysate. The protein content of each sample was determined using the BCA Protein Assay Kit (Thermo Scienti c). Then, equal amounts of proteins(12μg/lane) were separated on a 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidenedi uoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA).The membranes were blocked in 5% (w/v) nonfat dry milk in TBST (Tris-buffered saline-0.1% Tween) at 25 °C for 3 h and then incubated with the following primary antibodies: β-actin (1:800, Abcam, EPR16769), RAB35 (1:700, Abcam, ab152138), TSG101 (1:1500, Abcam, ab125011), CD63 (1:1000, Abcam, EPR21151), HSP70
(1:1000, Abcam, EPR16892). The bands were visualized using horseradish peroxidase (HRP) conjugated goat anti-rabbit IgG (1:2,000, Boster) prior to the ECL protocol (Amersham Biosciences, Piscataway, NJ, USA).
(1:1000, Abcam, EPR16892). The bands were visualized using horseradish peroxidase (HRP) conjugated goat anti-rabbit IgG (1:2,000, Boster) prior to the ECL protocol (Amersham Biosciences, Piscataway, NJ, USA).
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