Cat. # | Size | Qty. | Price |
---|---|---|---|
4125S | 100 µl |
|
REACTIVITY | H M R |
SENSITIVITY | Endogenous |
MW (kDa) | 86 |
SOURCE | Rabbit |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunofluorescence (Immunocytochemistry) | 1:100 |
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
Achieve higher quality immunofluorescent images using the efficient and cost-effective, pre-made reagents in our #12727 Immunofluorescence Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 24
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues adjacent to Ser183 of human TLK1. Antibodies are purified by protein A and peptide affinity chromatography.
Tousled-like kinases (TLK1 and TLK2) are nuclear serine/threonine kinases named for their homology to the Tousled gene from Arabidopsis thaliana, essential for flower development (1). The kinase activities of the TLKs are cell cycle regulated, with maximal activity during S phase (1). TLK appears to play a role in chromatin assembly and DNA damage checkpoint regulation (1,2). In C. elegans, TLK1 is essential for appropriate transcription during embryonic development (3). Substrates for TLK include the human chromatin assembly factor Asf, which functions in DNA replication- and repair-coupled chromatin assembly (2). DNA damage during S phase, when TLK is maximally active, leads to inhibition of TLK activity (1). This inhibition requires ataxia mutated kinase (ATM) and Chk1 (4,5). ATM and the related kinase ATR are activited by DNA damage during S phase, phosphorylate Chk1/Chk2, and block the transition into mitosis (6). Chk1 phosphorylates TLK1 on Ser743 in vitro and in vivo, leading to inhibition of TLK1 activity (4). This process likely provides a mechanism to slow the chromatin assembly processes controlled by TLK in the event of DNA damage.
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