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).
Alexa fluor 594 streptavidin
Manufactured by Yeasen
Sourced in China
Alexa-Fluor 594 Streptavidin is a fluorescent conjugate of the protein streptavidin. Streptavidin has a high affinity for the small molecule biotin, allowing the Alexa-Fluor 594 dye to be used as a detection or labeling reagent in various biological applications.
Automatically generated - may contain errors
2 protocols using alexa fluor 594 streptavidin
1
Immunohistochemical Analysis of Nav1.9 in DRG Neurons
2
Cell Proliferation Assay for HCC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HCC cells were seeded in five 96-well plates (1500 cells per well), and then the plates were removed on day 0, 1, 3, 5 and 7. Cells were treated with crystal violet buffer (0.1% w/v in methanol), stained cells were lysed in 10% acetic acid, and the 570 nm absorbance was detected or directly using cell counting Kit-8 (CCK8, beyotime, C0038) to measure 450nm absorbance according to the manufacturer’s construction. For EdU cell proliferation staining, HCC cells were plated in a 12-well U chamber (1×105 per well) (IBIDI, 81201) overnight. Subsequently, cells were subjected to 20 μM EdU treatment for 3 h, fixed with 4% paraformaldehyde and permeated with 0.1% Triton X-100. The cells were incubated with the click reaction mixture (10 μM biotin-azide, 0.5 mM CuSO4, 0.5 mM Tris (3-hydroxypropyltriazolylmethyl) amine (THPTA), 5 mM vitamin C and 5 mM aminoguanidine) for 2 h at room temperature. After washing with PBS, the cells were stained with Alexa Fluor 488 Streptavidin (Biolegend, 405235) or Alexa Fluor 594 Streptavidin (Yeasen, 35107ES60) at room temperature for 1 h and then incubated with Hoechst 33342 for 10 min. Under a microscope magnified 200-fold, 5 random fields of view of each sample were selected to conduct the quantitative analysis of the percentage of cells in s-phase (EdU-positive cells/Hoechst-positive cells). Cells were counted using ImageJ software.
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!