|H Mk||Endogenous||47, 51||Rabbit|
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 and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
Annexin A7 Antibody detects endogenous levels of total annexin A7 protein, including isoforms 1 and 2.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human annexin A7. Antibodies are purified using protein A and peptide affinity chromatography.
Annexin A7/ANXA7 is a member of the annexin family of calcium/phospholipid-binding proteins, and is involved in the process of membrane fusion and exocytosis (1). Annexin A7 is a GTPase, and both GTP-binding and PKC activity are important in regulating protein function (2,3). Membrane binding of annexin A7 is calcium dependent (4). Two isoforms exist due to alternative splicing. Subcellular localization of annexin A7 has been shown to be in the cytoplasm, vesicular structures, membrane and in adrenal chromaffin granules (5,6). Nuclear localization has been shown in the developing mouse central nervous system as well as in adult mouse brain (7). Annexin A7-deficient mouse studies show that the protein has a role in insulin secretion and calcium signaling (8) as well as cardiac intracellular calcium homeostasis electrical stability (9). The gene for annexin A7 is a putative tumor suppressor (10), and alterations in the copy number have been reported in prostate cancer (11). Annexin A7 expression has also been correlated with survival in human glioblastoma patients (12), and haploinsufficiency in mice may promote genetic instability leading to tumorigenesis (13).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. The Annexin V-FITC conjugate is a product manufactured for Cell Signaling Technology, Inc. by TAU Technologies BV.
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