Sixty-two formalin-fixed paraffin-embedded (FFPE) tissues from bladder cancer patients of various stages and grades were collected from Hospital Kuala Lumpur. The cohort of tissues comprised of 17 Grade 1, 18 Grade 2, and 27 Grade 3 tumours, of which, there were 18 pTa, 19 pT1, 11 pT2, and 14 pT3 and pT4 stages. Tissues were deparaffinized at 36°C for 1 hour. Slides were sequentially rehydrated with 100%, 95%, 80%, and 70% ethanol immersions, followed by antigen retrieval which was done by boiling the slides in sodium citrate using a microwave. The sections were then incubated with a primary polyclonal rabbit antibody (ab85567) against CERS2 (Abcam, Cambridge, United Kingdom) at 4°C overnight and then incubated with Dako DAB chromogen (Dako, Glostrup, Denmark) secondary antibody at room temperature for 10 minutes. A negative control was run in every experiment by replacing the primary antibody with 10% TBS. Sections were finally counterstained using haematoxylin and sequentially dehydrated with 70%, 80%, 95%, and 100% ethanol immersions, mounted with DPX, and then observed under the microscope. Scoring was based on staining intensity.
Ab85567
Manufactured by Abcam
Sourced in United Kingdom
Ab85567 is an Antibody that can be used for laboratory research purposes. It is a product manufactured by Abcam.
Automatically generated - may contain errors
2 protocols using ab85567
1
CERS2 Expression in Bladder Cancer
2
Schwann Cell Protein Expression Analysis
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After co-culturing for 72 h, the Schwann cells from the bottom layer were collected. The protein concentration was determined by a bicinchoninic acid protein assay kit (Beyotime; Nantong, China) in accordance with the manufacturer’s protocol. Equal amounts of protein extracts were applied to 10% sodium dodecyl sulfate-polyacrylamide gels at 80 V for 20 min, then 120 V for 50 min, followed by a transfer to a polyvinylidene difluoride membrane at 15 V for 30 min. The membrane was incubated with rabbit polyclonal antibody against anti-NRG1 antibody (Abcam Cat# ab53104, RRID:AB_881750; Cambridge, UK), anti-GFAP antibody (Abcam Cat# ab7260, RRID:AB_305808; Cambridge, UK), anti-Ki67 antibody (Abcam Cat# ab16667, RRID:AB_302459; Cambridge, UK), or mouse antibody against GAPDH (Abcam Cat# ab8245, RRID:AB_2107448; Cambridge, UK) and anti-Cers2 antibody (Abcam Cat# ab85567, RRID:AB_1925170; Cambridge, UK). The protein bands were exposed using a scanner.
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