|H M R||Endogenous||78||Rabbit IgG|
Western blot analysis of extracts from COS-7 cells, mock transfected (-) or transfected with constructs expressing full-length human PKG-1α (hPKG-1α; +) or PKG-1β (hPKG-1β; +), using PKG-1α (D10G2) Rabbit mAb (upper) or PKG-1 (C8A4) Rabbit mAb #3248 (lower).Learn more about how we get our images.
Western blot analysis of extracts from 293T and SH-SY5Y cells using PKG-1α (D10G2) 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.
PKG-1α (D10G2) Rabbit mAb recognizes endogenous levels of total PKG-1α protein. This antibody does not cross-react with PKG-1β.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human PKG-1α protein.
Cyclic GMP-dependent kinases (cGK/PKG) belong to the AGC family of serine/threonine protein kinases. In mammals, two genes encode PKG-1 and PKG-2. Alternative PKG-1 splicing yields α and β isoforms, which display tissue-specific expression patterns in humans (1). All PKG family members are activated by increased cellular cGMP, which binds to the enzyme's regulatory domain inducing a conformational change and leading to enzyme activation. cGMP levels are increased through the activation of guanylyl cyclases, a process known to occur in part through nitric oxide (NO) signaling (2).
In addition to well established roles in platelet activation and smooth muscle relaxation (3), PKG signaling is important in many biological processes including cardiac contractility, axon guidance, bone growth, contraction of intestinal smooth muscle, and erectile dysfunction (4).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|13511S||100 µl (10 western blots)||$ 255.0|