The total hCSC proteins were extracted and the protein concentrations were detected using a BCA Protein Assay Kit (Solarbao, China). The protein samples were run through sodium dodecylsulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and transferred to polyvinylidene fluoride (PVDF) films. After blocking, the membranes were incubated with antibodies, including rabbit anti‐ALDH3A1(1:500, Abcam), rabbit anti‐keratocan (1:100, Abcam), rabbit anti‐lumican (1:1000, Abcam), rabbit anti‐fibronectin (1:1000, Abcam), rabbit anti‐α‐SMA (1:1000, Abcam) and mouse anti‐GAPDH overnight at 4℃. The membranes were then incubated with anti‐mouse or anti‐rabbit IRDye 680RD secondary antibodies (1:10 000, LI‐CORBiosciences) for 2 h at room temperature. Bands were visualized with the Odyssey Fc Imaging System (LI‐COR Biosciences, USA) and quantified with Image J software. The expression ratios of the target proteins were determined after normalizing the individual GADPH levels. The results from three independent experiments were statistically analysed.
Rabbit anti lumican
Manufactured by Abcam
Sourced in United Kingdom
Rabbit anti-lumican is a primary antibody that recognizes the lumican protein. Lumican is a small leucine-rich proteoglycan involved in the regulation of collagen fibril assembly. This antibody can be used for the detection and quantification of lumican in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti fibronectin, by Abcam (4 mentions)
Lsm800 confocal microscope, by Zeiss (4 mentions)
Rabbit anti aldh3a1, by Abcam (3 mentions)
Rabbit anti α sma, by Abcam (3 mentions)
Rabbit anti keratocan, by Abcam (2 mentions)
Irdye 680rd secondary antibody, by LI COR (2 mentions)
Odyssey fc imaging system, by LI COR (2 mentions)
Rabbit anti nanog, by Abcam (1 mentions)
Rabbit anti klf4, by Abcam (1 mentions)
Rabbit anti pax6, by Abcam (1 mentions)
Mouse anti abcg2, by Abcam (1 mentions)
Rabbit anti sox2, by Abcam (1 mentions)
4 protocols using rabbit anti lumican
1
Quantitative Protein Analysis in hCSCs
2
Western Blot Analysis of Eye Proteins
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Total proteins were extracted, and the protein concentrations were detected using a BCA Protein Assay Kit (Solarbao, China). The protein samples were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membranes. After blocking, the membranes were incubated with antibodies, including rabbit anti-ALDH3A1 (1:500, Abcam, United Kingdom), rabbit anti-keratocan (1:100, Abcam, United Kingdom), rabbit anti-lumican (1:1,000, Abcam,VUK), rabbit anti-fibronectin (1:1,000, Abcam, United Kingdom) and mouse anti-GAPDH overnight at 4°C. The membranes were then incubated with anti-mouse or anti-rabbit IRDye 680RD secondary antibodies (1:10,000, LI-COR Biosciences, United States) for 1 h at room temperature. The protein bands were visualized with the Odyssey Fc Imaging System (LI-COR Biosciences, United States) and quantified with ImageJ software. The expression ratios of the target proteins were determined after normalizing to the GAPDH levels. The results from three independent experiments were statistically analyzed.
3
Immunolabeling of Cellular Markers
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After fixation, permeabilizing and blocking, cells at P4 were then incubated overnight at 4℃ with primary antibodies, including rabbit anti‐ALDH3A1(1:200, Abcam), rabbit anti‐lumican (1:400, Abcam), rabbit anti‐Fibronectin (1:200, Abcam) and rabbit anti‐α‐SMA (1:500, Abcam) followed by washing three times in PBS and incubation with FITC‐conjugated anti‐rabbit and Cy3‐conjugated anti‐rabbit IgG secondary antibodies (1:500, Life) for 1 h at room temperature and were incubated with DAPI for nuclear staining for 15 min. Finally, the imaging was performed using an LSM800 confocal microscope (Zeiss, Germany).
4
Immunofluorescence Profiling of Stem Cell Markers
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After the cells were fixed, permeabilized and blocked, they were incubated overnight at 4°C with primary antibodies, including rabbit anti-Lumican (1:500, Abcam, United Kingdom), rabbit anti-Fibronectin (1:200, Abcam, United Kingdom), rabbit anti-α-SMA (1:500, Abcam, United Kingdom), rabbit anti-Nanog (1:100, Abcam, United Kingdom), rabbit anti-Sox2 (1:250, Abcam, United Kingdom), mouse anti-Oct4 (1:100, CST, United States), rabbit anti-Klf4 (1:100, Abcam, United Kingdom), mouse anti-ABCG2 (1:50, Abcam, United Kingdom), rabbit anti-Pax6 (1:50, Abcam, United Kingdom), rabbit anti-Myosin Ⅱa (1:50, CST, United States), rabbit anti-HIF1α (1:800, CST, United States) and rabbit anti-YAP1 (1:200, Bioss, China) followed by washing three times in PBS and incubation with FITC-conjugated anti-rabbit or anti-mouse IgG secondary antibodies (1:500, Life, United States) for 1 h at room temperature. Cells were then incubated with DAPI for nuclear staining for 15 min. Finally, imaging was performed using an LSM800 confocal microscope (Zeiss, Germany). The fluorescence intensity was quantified using ImageJ software (version 1.47, National Institutes of Health, United States).
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