After treatments, kidney slides and podocyte culture coverslips (cell density is 1000/well in 24-well-plate) were fixed, blocked and incubated with primary antibodies against NLRP3 (1:100, Abcam, Cambridge, MA, USA), cleaved-caspase-1 (1:200, Santa Cruz Biotechnology, Dallas, TX, USA), VPS16 (1:100, Proteintech Group, Chicago, IL, USA), IL-1β (1:200, R&D systems, USA), Lamp-1(1:100, Abcam, Cambridge, MA, USA) at 4 °C overnight. Then samples were incubated with corresponding secondary antibodies with either Alexa- 488- or Alexa-555-labeled (Invitrogen) for 1 h at room temperature in the dark room. Finally, samples were mounted with mounting medium containing DAPI sealed with nail polish and pictures were taken under confocal laser scanning microscope (Fluoview FV1000; Olympus, Tokyo, Japan). There were 5–6 slides in every group, and 3 to 5 frames were chosen in every sample at random to show the characteristics of cell statues. Co-localization coefficient was analyzed with Image Pro Plus 6.0 software (Media Cybernetics, Bethesda, MD) and expressed as Pearson’s correlation coefficient (PCC) [26 (link)].
Vps16
Manufactured by Proteintech
Sourced in United States
The VPS16 is a laboratory equipment designed for protein purification. It features a vacuum-driven system for efficient separation and concentration of protein samples. The VPS16 allows for consistent and reliable protein recovery, making it a valuable tool for researchers and scientists working in the field of protein biochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Cleaved caspase 1, by Santa Cruz Biotechnology (2 mentions)
Image pro plus 6, by Media Cybernetics (2 mentions)
Alexa 488, by Thermo Fisher Scientific (2 mentions)
Fluoview fv1000, by Olympus (2 mentions)
Lamp1, by Abcam (2 mentions)
Il 1β, by R&D Systems (2 mentions)
Nlrp3, by Abcam (2 mentions)
Sqstm1 p62, by Abcam (1 mentions)
Cathepsin d, by Merck Group (1 mentions)
Quantity one, by Bio-Rad (1 mentions)
Lamp2, by Merck Group (1 mentions)
Gapdh, by Proteintech (1 mentions)
3 protocols using vps16
1
Immunofluorescence Analysis of Podocyte Markers
2
Western Blot Analysis of Autophagy Markers in Hippocampal CA1 Region
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rats were killed under chloral hydrate anesthesia at 0, 4, 24 and 48 h after reperfusion with or without hypoxia (n=5 in each group). The brain tissue was cut into 2 mm coronal slices using a brain matrix and the CA1 subregions of bilateral hippocampi were quickly dissected under a stereomicroscope. Proteins of the hippocampal CA1 subregion were extracted as described previously.2 (link) Western blot was performed as described previously2 (link) using the antibodies including LC3 (1:10000; Novus), Atg5 (1:1000; Abcam), SQSTM1/p62 (1:2000; Abcam), LAMP2 (1:6000; Sigma), Cathepsin D (1:1000; Sigma), Rab7 (1:1000; Abcam), Vps16 (1:1000; Proteintech Group, Inc. Chicago, IL, USA), UVRAG (1:1000; Millipore) and GAPDH (1:10000; Proteintech). Densitometric analysis for the quantification of the bands was performed using image analysis software (Quantity One, Bio-Rad Laboratories, Inc., Hercules, CA, USA). Relative optical densities of protein bands were calibrated with β-actin or GAPDH and normalized to those in Sham-operated rats.
3
Immunofluorescence Analysis of Podocyte Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After treatments, kidney slides and podocyte culture coverslips (cell density is 1000/well in 24-well-plate) were fixed, blocked and incubated with primary antibodies against NLRP3 (1:100, Abcam, Cambridge, MA, USA), cleaved-caspase-1 (1:200, Santa Cruz Biotechnology, Dallas, TX, USA), VPS16 (1:100, Proteintech Group, Chicago, IL, USA), IL-1β (1:200, R&D systems, USA), Lamp-1(1:100, Abcam, Cambridge, MA, USA) at 4 °C overnight. Then samples were incubated with corresponding secondary antibodies with either Alexa- 488- or Alexa-555-labeled (Invitrogen) for 1 h at room temperature in the dark room. Finally, samples were mounted with mounting medium containing DAPI sealed with nail polish and pictures were taken under confocal laser scanning microscope (Fluoview FV1000; Olympus, Tokyo, Japan). There were 5–6 slides in every group, and 3 to 5 frames were chosen in every sample at random to show the characteristics of cell statues. Co-localization coefficient was analyzed with Image Pro Plus 6.0 software (Media Cybernetics, Bethesda, MD) and expressed as Pearson’s correlation coefficient (PCC) [26 (link)].
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!