We have developed a protocol for rapid isolation of cytokines and signaling molecules from intact tissue. This method is for total protein extraction and makes use of a non-abrasive tissue extraction reagent. Tissue samples as little as 10 mg may be e...
Thermo Scientific 1-Step human in vitro translation (IVT) kits use the coupled format and are optimized for protein expression from DNA templates. However, protocols for using these same reagents for mRNA (translation only) are av...
There is no universal protocol for protein sample preparation. Sample preparation protocols must take into account several factors, such as the source of the specimen or sample type, chemical and structural heterogeneity of proteins, the cellular or subcellular location of the protein of interest, the...
Cell-free protein synthesis is a versatile protein production system. Performance of the protein synthesis depends on highly active cytoplasmic extracts. Extracts fromE. coliare believed to work best; they are routinely obtained from exponential growing cells, aiming to capture the most active translatio...
alkyne cycloaddition (CuAAC) and alkyne agarose-based strategy for scalability and high reaction rates32as the basis for protocol optimization. We individually evaluated key experimental steps to improve protein extraction from tissue and click chemistry efficiency by systematic implementation of previous ...
The tissue labeling protocol was followed as previously reported in the protocol35, including antigen retrieval, protein blocking, metal-tagged antibody labeling, and nucleus counterstains. The protocol starts with baking the FFPE sections at 60 oC for 2 hours to ensure that the tissues adhere ...
Protocol Springer Protocols(2024) Neural Stem/Progenitor Cell (NSPC) Extraction and Culture Jemma Gasperoni,Sebastian Dworkin Neural stem–progenitor cells (NSPCs) are multipotent, self-renewing cells that generate radial glial cells (RGC). RGCs then give rise to neurons and glia during neural deve...
For example, Patch-seq10 is a protocol for collecting RNA, electrophysiological (ephys), and morphological properties from the same neuron, and is critical for linking the molecular and cellular properties of neurons. Patch-seq navigates a micropipette to a neuron of interest in order to ...
We found that knockdown of genes related to LPS core and O-antigen synthesis and protein glycosylation did not significantly impair outer membrane integrity, except for hldD (Fig. 3B). Thus, increased antibiotic susceptibility is not simply due to increased antibiotic influx alone....
Cells were then re-suspended in PBS (with 0.05% bovine serum albumin) and subjected to cell loading, microbead loading, and cell lysis as described in the Well-paired-seq protocol. Briefly, single cells were loaded into the lower microwells by gravity, and cell clusters and cell-free RNAs ...