|H||Transfected Only||50||Rabbit IgG|
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing DDK/Myc-tagged full-length human Lmx1B protein (hLmx1B-DDK/Myc; +), using Lmx1B (D1E2) Rabbit mAb.Learn more about how we get our images
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 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: 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
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.
Lmx1B (D1E2) Rabbit mAb recognizes transfected levels of total Lmx1B protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human Lmx1B protein.
LIM homeobox transcription factor 1 β (Lmx1B) is a member of an evolutionarily conserved family of transcription factors that regulate developmental pattern formation in both vertebrates and invertebrates (1). Numerous developmental studies show that Lmx1B is required for vertebrate dorsoventral limb patterning, as well as normal glomerular basement membrane development and typical differentiation of central serotonergic neurons (2,3).
Mutations in the corresponding Lmx1B gene have been associated with nail-patella syndrome (NPS), an autosomal dominant disorder characterized by dysplasia of fingernails, skeletal anomalies and, frequently, renal disease (2). Abnormal developmental disorders such as developmental glaucoma and idiopathic Parkinson’s disease have also been associated with Lmx1B function (4,5).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|13457S||100 µl (10 western blots)||$255.00.0|