Resources results for "FBL"

WW Protein Domain

A scientific resource for the WW protein domain containing information on structure, function, and domain binding to proline-rich motifs.

Resources

Our protocol is as follows:

REAGENTS:
TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174
Lipopolysaccharides (LPS) #14011

DAY 1:

1. Seed THP-1 cells at approximately 8 X 10^5 cells/mL in 15 cm plates with 20 ml RPMI +10% FBS + 0.05 mM 2-mercaptoethanol. (We use plates instead of flasks since the cells adhere to the plate after treatment with TPA).
2. Treat plates overnight with 80nM TPA.

1. Wash treated plates with 10 mL 1x PBS, drain, and add fresh media (RPMI +10% FBS + 0.05 mM 2-mercaptoethanol).
2. Allow cells to rest overnight.

Resources

We use Microcon columns from Amicon (30 kDa cut off) to concentrate media. It is important that the medium does not contain FBS because this will make it difficult to concentrate with the Microcon columns.
1. Spin the media (20 ml) to pellet any cellular debris.
2. Add the 20ml volume to a Microcon column and concentrate it down to 500 ul.
3. Add 10 ml of cold 1xPBS to the 500 ul sample to wash it and concentrate again to 500 ul.
4. Transfer the 500 ul of concentrated media to an eppendorf tube and wash the Microcon tube membrane twice with 250 ul of cold 1xPBS to remove any proteins stuck to the membrane. This should result in a 1ml sample.
5. Add 500 ul of 3x reducing SDS sample buffer to 1.0 ml of concentrated media. MMPs tend to be quite unstable and degrade easily in our hands, so we highly suggest adding protease and phosphatase inhibitors to the lysate, along with PMSF and 5 mM EDTA (as a chelator).
6. Aliquot the lysates and store them at -80C.

Hallmarks of Antibody Validation: Multiple Antibody Strategy

A multiple-antibody strategy is a powerful approach to antibody validation. The most common method to achieve this is to immunoprecipitate (IP) the target with one antibody and subsequently detect it by western blotting with another antibody against the

CUT&Tag: An Introduction

Learn how CUT&Tag cuts down your chromatin profiling experiment time. You’ll get the same great data with a fraction of the DNA library prep time and improved signal-to-noise.

CAR-T Therapy Research Tools: Discover Efficacious Therapies Faster

Learn about the antibodies, custom reagents, and ready-to-use cell assay kits to support your CAR-T therapy development.

Histone Modification Table

The Histone Modification Table provides a referenced list of many known histone modifications, associated modifying enzymes, and proposed functions.