Western blot analysis of extracts from K-562 and LNCaP cells using TXNIP (D5F3E) Rabbit mAb (Biotinylated).
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This Cell Signaling Technology antibody is conjugated to biotin under optimal conditions. The biotinylated antibody is expected to exhibit the same species cross-reactivity as the unconjugated TXNIP (D5F3E) Rabbit mAb #14715.
Supplied in 136 mM NaCl, 2.6 mM KCI, 12 mM sodium phosphate (pH 7.4) dibasic, 2 mg/ml BSA, and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
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.
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.
Do not add Anti-biotin, HRP-linked Antibody for detection of biotinylated protein markers. There is no need. The Streptavidin-HRP will also visualize the biotinylated markers.
* 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 June 2016
Protocol Id: 266
TXNIP (D5F3E) Rabbit mAb (Biotinylated) recognizes endogenous levels of total TXNIP protein.Species Reactivity:
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val337 of human TXNIP protein.
The ubiquitously expressed thioredoxin-interacting protein (TXNIP) binds and inhibits thioredoxin to regulate cellular redox state (1-3). Research studies demonstrate that hyperglycemia induces TXNIP expression and increases cellular oxidative stress (1). In addition, these studies show that TXNIP reduces glucose uptake directly by binding the glucose transporter Glut1 to stimulate receptor internalization or indirectly by reducing Glut1 mRNA levels (3). Additional studies indicate that TXNIP plays a role in the regulation of insulin mRNA transcription (4). Microarray analyses indicate that TXNIP acts downstream of PPARγ and is a putative tumor suppressor that may control thyroid cancer cell progression (5). In addition, the TXNIP protein may be a potential therapeutic target for the treatment of type 2 diabetes and some disorders related to ER-stress (6).
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