50 μm sections from OCT frozen kidney cortical specimen were fixed overnight with 4% paraformaldehyde or 50 μm sections were cut by vibratome from 4% paraformaldehyde fixed cortical tissue fragments underwent staining with DAPI and antibodies for the specified markers including: Megalin, LRP2 (cat#ab76969,abcam), Aquaporin-1 (AQP1,clone 1/22, cat# sc-32737, Santa Cruz Biotechnology), Slc12A3 (cat#HPA028748, Sigma), Uromodulin, Tamm-Horsfall protein (cat# AF5144, R&D Systems), CD45 (clone HI30,cat# 304002), CD31 (clone JC/70A cat# NB600–562-R, Novus), Nestin (clone 10C2, cat# 656810, Biolegend), and Cytokeratin-8 (cat# NBP-34267, Novus). All staining was performed in 1X Phosphate buffer saline with 10% normal goat serum and 0.1% Triton X-100 with overnight incubations. All secondaries were raised in goat highly cross-adsorbed and were labeled with Alexa488, Alexa546, Alexa555, or Alexa568, Alexa647. The CD31, CD45, and Nestin antibodies were directly conjugated with Dylight 550, Alexa 488, and Alexa 647, respectively. Stained tissue was mounted under #1.5 coverslips in Prolong Glass allowed to cure for 24–48 h before sealing with nail-polish. Mounted and sealed tissue was stored at 4°C before imaging.
Nestin clone 10c2
Manufactured by BioLegend
Nestin (clone 10C2) is a mouse monoclonal antibody that recognizes the intermediate filament protein Nestin. Nestin is a marker for neural stem and progenitor cells.
Automatically generated - may contain errors
2 protocols using nestin clone 10c2
1
Immunofluorescence Profiling of Kidney Cortex
2
Comprehensive Kidney Tissue Immunostaining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Three deceased donor kidney nephrectomy tissue were used based on a protocol approved by the Institutional Review Board at Indiana University. 50 µm section from OCT frozen kidney cortical specimen were fixed overnight with 4% paraformaldehyde or 50 µm sections were cut by vibratome from 4% paraformaldehyde fixed cortical tissue fragments underwent staining with DAPI and antibodies for the specified markers including: Megalin, LRP2 (cat#ab76969 ,abcam), Aquaporin-1 (AQP1,clone 1/22, cat# sc-32737, Santa Cruz Biotechnology), Slc12A3 (cat#HPA028748, Sigma), Uromodulin, Tamm-Horsfall protein (cat# AF5144, R&D Systems), CD45 (clone HI30,cat# 304002,), CD31 (clone JC/70A cat# NB600-562-R, Novus), Nestin (clone 10C2, cat# 656810, Biolegend) and Cytokeratin-8 (cat# NBP-34267, Novus). All staining was performed in 1X Phosphate buffer saline with 10% normal goat serum and 0.1% Triton X-100 with overnight incubations. All secondaries were raised in goat highly crossadsorbed, and were labeled with Alexa488, Alexa546, Alexa555 or Alexa568, Alexa647. The CD31, CD45 and Nestin antibodies were directly conjugated with Dylight 550, Alexa488 and Alexa 647, respectively. Stained tissue was mounted under #1.5 coverslips in Prolong Glass allowed to cure for 24-48 hours before sealing with nail-polish. Mounted and sealed tissue was stored at 4C before imaging.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!