|H M R||Endogenous||40-48||Rabbit IgG|
Western blot analysis of extracts from various cell lines using Tristetraprolin (D1I3T) Rabbit mAb. KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.Learn more about how we get our images
Western blot analysis of extracts from Raw 264.7 cells treated with Lipopolysaccharides (LPS) #14011 (1 μg/ml, 4 hr), with (+) or without (-) treatment with phophatases, Calf Intestinal Phophatase (CIP) and λ-Phosphatase (λ-Phosphatase).Learn more about how we get our images
Western blot analysis of extracts from THP-1 cells, differentiated with TPA #4174 (80 nM, 24 hr) followed by treatment with Lipopolysaccharides (LPS) #14011 (1 μg/ml, 6 hr; +) or without additional treatment (-) using Tristetraprolin (D1I3T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).Learn more about how we get our images
Western blot analysis of extracts from Raw 264.7 cells, untreated (-) or treated with Lipopolysaccharides (LPS) #14011 (1 μg/ml, 4 hr; +) using Tristetraprolin (D1I3T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).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.
Tristetraprolin (D1I3T) Rabbit mAb recognizes endogenous levels of total Trisetraprolin protein.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala235 of human tristetraprolin protein.
Tristetraprolin (TTP), also known as NUP475, G0S24, RNF162A, TIS11, and ZFP36, is a CCCH tandem zinc-finger protein that binds to adenosine and uridine (AU)-rich elements (AREs) within 3'-untranslated regions of mRNA and leads to their rapid degradation (1-6). Expression of TTP is rapidly induced by mitogens and growth factors including insulin, phorbol ester, cytokines, and lipopolysaccharide (LPS). In addition, numerous phosphorylation sites on TTP can regulate its stability, nuclear to cytosolic trafficking, as well as controlling its ARE-binding activity. Many of the target mRNAs for TTP, such as TNF-α, have critical roles in inflammation and cancer (2), and mice deficient in TTP develop a systemic autoimmune inflammatory syndrome along with excessive TNF-α levels (7). Furthermore, suppression of TTP expression has been identified as a negative prognostic indicator for some cancers (8).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge. Tween is a registered trademark of ICI Americas, Inc.
Explore pathways related to this product.
|71632S||100 µl (10 western blots)||$255.00.0|