REACTIVITY | SENSITIVITY | MW (kDa) | Isotype |
---|---|---|---|
H M R Mk | Endogenous | 25 | Rabbit IgG |
Western blot analysis of extracts from various cell lines using NRMT (D9D6P) Rabbit mAb.
Learn more about how we get our imagesFor 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 datasheet or 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 (#13953, 5 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 November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
Application | Dilutions |
---|---|
Western Blotting | 1:1000 |
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
NRMT (D9D6P) Rabbit mAb recognizes endogenous levels of total NRMT protein.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu49 of human NRMT protein.
N-terminal RCC1 methyltransferase (NRMT), formerly known as methyltransferase-like protein 11A (METTL11A), is a member of the methyltransferase 11 family of proteins and is the first α-N-methyltransferase to be discovered in humans (1-3). Amino-terminal methylation of free α-amino groups is a post-translational modification where an initiating Met residue is cleaved and the exposed α–amino group is mono-, di-, or trimethylated by NRMT (4). NRMT methylates proteins containing an amino-terminal Met-X-Pro-Lys motif, where X is an alanine, proline, or serine residue (4). Substrates of NRMT include the Ran guanine nucleotide-exchange factor (RCC1), SET/TAF-1/PHAP-II, retinoblastoma (Rb), and CENP-B (3-6). α-N-methylation of RCC1 is required for efficient binding to chromatin, securing normal bipolar spindle formation and chromosome segregation (3,5). α-N-methylation of CENP-B also appears to regulate CENP-B binding to centromeric DNA (6).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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Product # | Size | Price |
---|---|---|
13432S | 100 µl (10 western blots) | $ 255.0 |