DRG neurons were collected from the lumbar spinal cord (L4–L5). The DRG were fixed in 4% PFA in PBS for 2 h 30 min at 4 °C then cryoprotected in PBS containing 30% sucrose overnight at 4 °C. The DRG were frozen in OCT, and cryosectioned at 12 μm. The sections were permeabilized in PBS containing 0.5% TritonX-100) for 10 min, and were blocked with 10% goat serum for 1 h. The sections were incubated for 24 h at 4 °C with polyclonal rabbit anti-Nav1.9 (1:200; alomone labs). Then the sections were incubated with biotinylated griffonia simplicifolia Lectin I isolectin B4 (10 μg ml−1, Vector Laboratories, California, USA) for 30 min at room temperature. After incubation, sections were washed three times for 5 min in PBST (0.05% Tween20). Finally, incubated with Alexa-Fluor 488-nm (1:500, Invitrogen) and Alexa-Fluor 594 Streptavidin (1:200, Yeasen Biotechnology, Shanghai, China). After being washed three times for 5 min in PBST, sections were mounted with coverslips. Fluorescence images were acquired with the FV1000 confocal microscope (Olympus, Tokyo, Japan).
Lectin 1 isolectin b4
Manufactured by Vector Laboratories
Sourced in United States
Lectin I isolectin B4 is a plant-derived protein that selectively binds to specific sugar residues on the surface of cells. It is commonly used in research applications to identify and visualize particular cell types or structures.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 594 streptavidin, by Yeasen (1 mentions)
Alexa fluor 488 nm, by Thermo Fisher Scientific (1 mentions)
Fv1000 confocal microscope, by Olympus (1 mentions)
α smooth muscle actin α sma, by Agilent Technologies (1 mentions)
Vs200 fluorescent slide scanner, by Olympus (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
2 protocols using lectin 1 isolectin b4
1
Immunohistochemical Analysis of Nav1.9 in DRG Neurons
2
Quantifying Cardiac Vascular and Myofibroblast Density
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorescein labeled Griffonia Simplicifolia Lectin I isolectin B4 (1:75; Vector Laboratories) was used to identify endothelial cells within the right ventricle, α-smooth muscle actin (α-SMA) (1:75; Dako) was used to identify smooth cells, and Hoechst 33342 (1:1000; Life Technologies) was used to identify nuclei. Slides were then mounted by using Fluoromount and scanned at 20× magnification using an Olympus VS200 fluorescent slide scanner. Arteriole and myofibroblast density was quantified by manually counting in 5 evenly distributed regions throughout the right ventricle in 6 slides (evenly spaced throughout the RV mid ventricular region) using Fiji. Arterioles were identified based on the following criteria: 1) co-staining of isolectin and α-SMA; 2) presence of a visible lumen; and 3) Feret diameter >10 μm as previously described.10 (link),14 (link) Capillaries were identified as percent positive area of isolectin not co-localized with α-SMA. Myofibroblasts were identified as cells staining positive for α-SMA and not found in vessels.15
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!