Cat. # | Size | Price | Inventory |
---|---|---|---|
91788S | 100 µl |
REACTIVITY | H M R |
SENSITIVITY | Endogenous |
MW (kDa) | 28, 29 |
SOURCE | Rabbit |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro245 of human PI31/PSMF1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Proteasome-binding protein PI31 (Proteasomal Inhibitor of 31kD) is encoded by the PSMF1 gene and regulates proteasome function in the cell, which is essential for cell function and survival. Regulated degradation of intracellular proteins is mediated by the ubiquitin-proteasome system (UPS) by a multi-step process (1,2). First, protein to be degraded is tagged with a chain of ubiquitin molecules. Ubiquitinated proteins are then cleaved into small peptides by the 26S proteasome. Proteasome activity by the UPS is regulated at additional levels, including transcriptional expression of proteasomal proteins, posttranslational modification, and by various regulatory proteins. PI31/PSMF1 is a structurally conserved proteasome regulator that binds to the 20S proteasome and functions to inhibit 20S proteasome hydrolysis of small peptides (3,4). Unexpectedly, PI31/PSMF1 promotes UPS-mediated protein breakdown in vivo, suggesting additional proteasome regulatory functions beyond inhibition of the 20S proteasome (5). PI31/PSMF1 also serves as an adaptor protein to transport proteasomes via cellular protein motors (6,7). This process may be particularly important in neurons where local proteasomal degradation at distal tips of neurons may play a critical role in the maintenance of neuronal architecture, circuitry, and function.
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