REACTIVITY | SENSITIVITY | MW (kDa) | Isotype |
---|---|---|---|
H M R | Endogenous | 38 | Rabbit IgG |
Western blot analysis of human, mouse, and rat brain membrane extracts using Na Channel β1 Subunit (D9T5B) Rabbit mAb.
Learn more about how we get our imagesImmunoprecipitation of sodium channel β1 subunit from mouse brain membrane extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Na Channel β1 Subunit (D9T5B) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Na Channel β1 Subunit (D9T5B) 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
This protocol is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
10X Cell Lysis Buffer: (#9803) To prepare 10 ml of 1X cell lysis buffer, add 1 ml cell lysis buffer to 9 ml dH2O, mix.
NOTE: Add 1 mM PMSF (#8553) immediately prior to use.
Proceed to one of the following specific set of steps.
NOTE: To minimize masking caused by denatured IgG heavy chains (~50 kDa), we recommend using Mouse Anti-Rabbit IgG (Light-Chain Specific) (L57A3) mAb (#3677) or Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127). To minimize masking caused by denatured IgG light chains (~25 kDa), we recommend using Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127).
posted December 2008
revised November 2013
Protocol Id: 409
Application | Dilutions |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:100 |
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.
Na Channel β1 Subunit (D9T5B) Rabbit mAb recognizes endogenous levels of total sodium channel β1 subunit protein. This antibody also cross-reacts with an unidentified protein of 60 kDa in whole brain lysate.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala51 of human sodium channel β1 subunit protein.
Mammalian voltage-gated sodium channels (VGSCs) are composed of a pore-forming α subunit and one or more regulatory β subunits (1). Four separate genes (SCN1B-SCN4B) encode the five mammalian β subunits β1, β1B, β2, β3, and β4. In general, β subunit proteins are type I transmembrane proteins, with the exception of secreted β1B protein (reviewed in 2). β subunits regulate α subunit gating and kinetics, which controls cell excitability (3,4). Sodium channel β subunits also function as Ig superfamily cell adhesion molecules that regulate cell adhesion and migration (5,6). Additional research reveals sequential processing of β subunit proteins by β-secretase (BACE1) and γ secretase, resulting in ectodomain shedding of β subunit and generation of an intracellular carboxy-terminal fragment (CTF). Generation of the CTF is thought to play a role in cell adhesion and migration (7,8). Multiple studies demonstrate a link between β subunit gene mutations and a number of disorders, including epilepsy, cardiac arrhythmia, multiple sclerosis, neuropsychiatric disorders, neuropathy, inflammatory pain, and cancer (9-13).
The sodium channel β1 subunit (SCN1B) plays a crucial role in neuronal migration and pathfinding during brain development (14). Mutations in the corresponding SCN1B gene are associated with generalized epilepsy with febrile seizures plus 1 (15), Brugada syndrome (16), and familial atrial fibrillation (17). A SCN1B loss of function mutation results in a severe form of pediatric epileptic encephalopathy known as Dravet syndrome (18).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. XP is a registered trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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Product # | Size | Price |
---|---|---|
14684S | 100 µl (10 western blots) | $ 255.0 |