|H M||Endogenous||90 to 95||Rabbit|
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using APS Antibody.Learn more about how we get our images.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using APS Antibody.Learn more about how we get our images.
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma, using APS Antibody.Learn more about how we get our images.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using APS Antibody in the presence of control peptide (left) or antigen specific peptide (right).Learn more about how we get our images.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, showing cytoplasmic localization using APS Antibody.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
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
posted June 2005
revised March 2016
Protocol Id: 306
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.
APS Antibody detects endogenous levels of total APS. This antibody does not cross-react with other adaptor/docking proteins.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino-terminus of human APS. Antibodies are purified by protein A and peptide affinity chromatography.
APS is an SH2 and PH domain-containing adaptor protein closely related to Lnk and SH2-B (1). APS was identified as a substrate for many receptor tyrosine kinases including TrkA, insulin receptor, c-Kit and PDGF receptor (2). Tyrosine phosphorylation of APS provides docking sites for downstrean signaling components, mediating diverse signaling pathways. APS plays quite different roles in RTK signaling. Overexpression of APS has been shown to inhibit PDGF-induced mitogenicity, which may result from APS/c-Cbl-mediated PDGF receptor degradation (3). However, APS promotes enhanced mitogenicity in response to insulin stimulation (4). The striking difference in APS-mediated signaling between the different RTKs could lie in the mode of interaction with the respective receptor.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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