Western blotting was carried out as previously described (8 (link)). Briefly, mTiD/PLiD lysates were prepared and quantified by using Bio-Rad protein assay kit (Thermo Scientific., Chicago, IL). Cell lysates were subjected to SDS-PAGE and blotted onto PVDF membrane (EMD Millipore, Danvers, MA), blocked and incubated with primary antibodies (mouse anti-E-cadherin (610181, BD Biosciences, San Jose, CA ), mouse anti-vimentin (sc-6260, Santa Cruz, Dallas, TX), rabbit anti-CD31 (PA5–16301, ThermoFisher Scientific, Grand Island, NY), rabbit anti-LYVE1 (ab14917, Abcam, Cambridge, MA), mouse anti-surfactant protein (SP-B, ab3282, or SP-D, ab1778 Abcam, Cambridge, MA), rabbit anti-carcinoembryonic antigen (CEA, ab135781, Abcam, Cambridge, MA), a mouse anti-MUC16 (CA125, ab10029, Abcam, Cambridge, MA) and mouse anti-GAPDH (sc-365062, Santa Cruz, Dallas, TX). Then incubated with appropriate sheep anti-mouse secondary antibodies (NA911V, GE healthcare Life sciences, Pittsburgh, PA) or donkey anti-rabbit (NA934V, GE healthcare Life sciences, Pittsburgh, PA) tagged with HRP. Proteins of interest were detected using enhanced chemiluminescence system (RPN2106, GE healthcare Life sciences, Pittsburgh, PA or 34095, ThermoFisher Scientific, Grand Island, NY) and imaged on chemiDoc™ XRS with Image lab software (Bio-Rad, Hercules, CA).
Ca125
Manufactured by Abcam
CA125 is a protein that is commonly found in the blood. It is often used as a biomarker for certain types of cancer, particularly ovarian cancer.
Automatically generated - may contain errors
Lab products found in correlation
Image lab software, by Bio-Rad (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
Na934v, by GE Healthcare (1 mentions)
Sc 6260, by Santa Cruz Biotechnology (1 mentions)
Mouse anti e cadherin, by BD (1 mentions)
Pa5 16301, by Thermo Fisher Scientific (1 mentions)
Ab14917, by Abcam (1 mentions)
Rpn2106, by GE Healthcare (1 mentions)
Sc 365062, by Santa Cruz Biotechnology (1 mentions)
Fibroblast growth factor (fgf), by Thermo Fisher Scientific (1 mentions)
Human chorionic gonadotropin (hcg), by Merck Group (1 mentions)
Vascular endothelial growth factor (vegf), by Merck Group (1 mentions)
Lactate dehydrogenase ldh, by Abcam (1 mentions)
Mirneasy serum plasma kit, by Qiagen (1 mentions)
Human xl oncology array kit, by R&D Systems (1 mentions)
Alkaline phosphatase alp, by Abcam (1 mentions)
2 protocols using ca125
1
Western Blotting for Cell Marker Analysis
2
Biomarker Screening for Teratoma Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Two milliliters of blood were collected directly after each MRI session via a tail vein catheter using EDTA-coated tubes. Blood was stored on ice for up to 3 hr before centrifugation at 3000 × g to separate the plasma. Plasma was aliquoted and stored at −80°C until specific ELISAs were performed. The following ELISA kits were acquired and performed following the manufacturer's protocols: CEA (Abcam), AFP (R&D Systems), FGF (Invitrogen), HCG (Sigma), vascular endothelial growth factor (Sigma), lactate dehydrogenase (LDH; Abcam), alkaline phosphatase (ALP; Abcam), and CA-125 (Abcam). For the exploratory biomarker study, plasma from a separate cohort of animals was used to screen for microRNAs or cancer-associated proteins. Total RNA, including miRNA, was extracted from plasma using an miRNeasy Serum/Plasma Kit (Qiagen) according to the manufacturer's protocol. Screening was performed using the Human Cancer Pathway Finder miRNA PCR array (Qiagen) according to the manufacturer's protocol. The human XL oncology array kit (R&D Systems) was used according to the manufacturer's protocol to screen for potential protein-based teratoma biomarkers.
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