Immunoblots of whole cell lysates were essentially as previously described (19 (link)). Primary antibodies were as follows: CK5 (mouse NCL-L-CK5, Leica Biosystems, Buffalo Grove, IL, USA, 1:1500), E-cadherin (mouse, 14472, Cell Signaling Technologies, 1:1000), CK17 (mouse, NBP2–29421, Novus Biologicals, Littleton, CO, 1:1000), CK8/18 (mouse, NCL-L-5D3, Leica Biosystems, 1:2000), Vimentin (rabbit, 5741, Cell Signaling Technologies, 1:1000), ZEB1 (rabbit, NBP1–05987, Novus Biologicals, 1:1000) or β-actin (mouse, A5441, Sigma, 1:1000). Secondary antibodies were IRDye800CW Goat-Anti-Mouse IgG (926-32210, Li-Cor Biosciences, Lincoln, NE, USA) and IRDye 680LT Goat-Anti-Rabbit IgG (926-68021, Li-Cor Biosciences) both at 1:10,000. Immunoblots were imaged and analyzed with the Odyssey Infrared Imaging System and Image Studio Lite (Li-Cor Biosciences).
Ncl l 5d3
Manufactured by Leica
Sourced in United States
The NCL-L-5D3 is a laboratory equipment product. It is designed for specific functions within a laboratory setting. No further details can be provided in an unbiased and factual manner without risk of extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Ncl l ck5, by Leica (2 mentions)
Irdye 680lt goat anti rabbit igg, by LI COR (1 mentions)
Image studio lite, by LI COR (1 mentions)
Irdye 800cw goat anti mouse igg, by LI COR (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
Pr1294, by Agilent Technologies (1 mentions)
Aperio digital pathology system, by Leica (1 mentions)
Impact dab peroxidase substrate kit, by Vector Laboratories (1 mentions)
Anti mouse ig mp 7402, by Vector Laboratories (1 mentions)
Imagescope software, by Leica (1 mentions)
2 protocols using ncl l 5d3
1
Immunoblot Analysis of Cell Lysates
2
Immunohistochemistry Protocol for Breast Cancer Markers
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IHC was performed as previously described.28 (link) Briefly, 10 μm paraffin sections were subjected to antigen retrieval in citrate buffer. Primary antibodies used were as follows: ERα (SP1, ThermoFisher, 1:100), PR (1294, DAKO, 1:500), CK5 (NCL-L-CK5, Leica Biosystems, 1:500), CK8/18 (NCL-L-5D3, Leica Biosystems, 1:500), and CA2 (SAB2700301, Sigma, 1:1600). Secondary antibodies were anti-Rabbit Ig MP-7401 (Vector, 1:2000; Burlingame, CA, USA) or anti-Mouse Ig MP-7402 (Vector, 1:2000). Development used the ImPACT DAB Peroxidase Substrate Kit (Vector). Images were captured using the Aperio Digital Pathology system (Leica Biosystems) and assembled in Adobe Photoshop CC with minimal linear changes to contrast. Quantification of the percentage of positive cells was performed using the Imagescope software (Leica) that used algorithms tuned for nuclear or cytoplasmic staining.
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