特异性ab12108 detects two bands >100kDa in WB. The higher MW band is probably a phospho protein as the signal is decreased when the blot is incubated with phosphatase prior to ab12108 (data not shown). However, this upper band is not GluR1 protein, as it is only detected by ab12108 and not by other GluR1 antibodies; it is therefore an unknown protein which is bound to specifically by ab12108.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml.
Use a concentration of 0.3 - 1 µg/ml. Predicted molecular weight: 102 kDa.
功能Ionotropic glutamate receptor. L-glutamate acts as an excitatory neurotransmitter at many synapses in the central nervous system. Binding of the excitatory neurotransmitter L-glutamate induces a conformation change, leading to the opening of the cation channel, and thereby converts the chemical signal to an electrical impulse. The receptor then desensitizes rapidly and enters a transient inactive state, characterized by the presence of bound agonist.
组织特异性Widely expressed in brain.
序列相似性Belongs to the glutamate-gated ion channel (TC 1.A.10.1) family. GRIA1 subfamily.
翻译后修饰Palmitoylated. Depalmitoylated upon glutamate stimulation. Cys-603 palmitoylation leads to Golgi retention and decreased cell surface expression. In contrast, Cys-829 palmitoylation does not affect cell surface expression but regulates stimulation-dependent endocytosis.
Western blot - Anti-Glutamate Receptor 1 (AMPA subtype) (phospho T840) antibody (ab12108)This image is courtesy of Chris Anderson, Wellcome Trust Sanger Institute, United Kingdom
Lanes 1, 4, 7 & 8 : Anti-Glutamate Receptor 1 (AMPA subtype) (phospho T840) antibody (ab12108) at 3 µg/ml Lane 2 : Lane 3 : Preimmune mouse IgG antibody Lane 5 : Blot reprobed with GluR1 mouse antibody Lane 6 :
Lane 1 : Mouse brain extract that has not been through IP protocol Lane 2 : Blank Lane 3 : Control IP for mouse IgG Lane 4 : Mouse brain extract following IP for GluR1 Lane 5 : As above Lane 6 : Blank Lane 7 : Input lane reprobed with GluR1 mouse antibody Lane 8 : IP lane reprobed with GluR1 mouse antibody
This image is courtesy of Chris Anderson, Wellcome Trust Sanger Institute, United Kingdom
Lane 1 (input) has two bands >100kDa as seen in the WB for ab12108, this lane is loaded with brain extract that has not been through the IP protocol. The higher MW band in the Lane 1 (input) is probably a phospho protein as the signal is decreased when the blot is incubated with phosphatase prior to ab12108 (data not shown). However, this upper band is not GluR1 protein, as it is only detected by ab12108 and not by other GluR1 antibodies; it is therefore an unknown protein which is bound to specifically by ab12108.
Lane 3 is a control IP with mouse IgG - this is the result of using a 'pre-immune' mouse IgG antibody to see if anything is pulled down from the brain extract independent of the GluR1 antibody. The ~55kDa bands in this lane correspond to IgGs.
Lane 4 is a WB for ab12108 following IP for GluR1. This lane should only contain GluR1 (band at ~100kDa) and anything bound to it (plus IgG). The single band at the right size for GluR1 in Lane 4 is therefore likely
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Glutamate Receptor 1 (AMPA subtype) (phospho T840) antibody (ab12108)This image is courtesy of an abreview submitted by Sophie Pezet, CNRS, Paris, France
Immunohistochemistical detection of Glutamate Receptor 1 (AMPA subtype) (phospho T840) using antibody ab12108 on PFA perfusion-fixed free floating rat brain sections. Primary Antibody dilution 1/100 incubated for 18 hours @ 20°C in PBS + 0.3 % Triton X100. Secondary Antibody: Goat anti-rabbit Alexa Fluor® 488 (1/1000). The antibody produced some staining in cytoplasmic neuronal inclusions. The picture shows the staining obtained using the objective X40 at the level of the cerebral cortex. The tissues were perfusion fixed with 4% PFA and later postfixed overnight in the same fixative. They were cryoprotected in 30% sucrose and cut using a cryostat.
Guzman RE et al. Involvement of ClC-3 chloride/proton exchangers in controlling glutamatergic synaptic strength in cultured hippocampal neurons. Front Cell Neurosci8:143 (2014).
Read more (PubMed: 24904288) »
Gray EE et al. Inhibitory interactions between phosphorylation sites in the C terminus of a-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type glutamate receptor GluA1 subunits. J Biol Chem289:14600-11 (2014).
Read more (PubMed: 24706758) »