SpecificityDetects protein interacting with Protein Kinase C (PICK 1). By Western blot, this antibody detects an ~52 kDa protein representing PICK 1 from AtT20 cell extract. Immunocytochemical staining of CV1 cells transfected with PICK 1 results in a diffuse staining pattern when expressed alone and coclustered when coexpressed with mGluR7.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1.7 µg/ml.
Use at an assay dependent concentration.
Use a concentration of 2 µg/ml. Detects a band of approximately 52 kDa.Can be blocked with PICK1 peptide (ab4964).
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
FunctionProbable adapter protein that bind to and organize the subcellular localization of a variety of membrane proteins containing some PDZ recognition sequence. Involved in the clustering of various receptors, possibly by acting at the receptor internalization level. Plays a role in synaptic plasticity by regulating the trafficking and internalization of AMPA receptors. May be regulated upon PRKCA activation. May regulate heteromeric ACCN3/ACCN2 channel.
Immunocytochemistry/ Immunofluorescence - Anti-PICK1 antibody (ab3420)Image courtesy of anonymous Abreview
ab3420 staining PICK1 in mouse Hippocampal neurons by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde and permeabilized in 0.1% saponin/PBS prior to blocking in 3% BSA for 1 hour at room temperature. The primary antibody was diluted 1/300 in 3% BSA, 0.1% saponin /PBS and incubated with the sample for 18 hours at 4 °C. The secondary antibody was Alexa Fluor® 455-conjugated Donkey anti-Rabbit polyclonal, diluted 1/1000.
Western blot - PICK1 antibody (ab3420)This image is courtesy of an anonymous Abreview
Lane 1 : An myc antibody in PBST for 16 hours at 4°C Lane 2 : Anti-PICK1 antibody (ab3420) at 1/500 dilution (in PBST for 16 hours at 4°C)
Lane 1 : Transfected CHO cells expressing human Pick1 tagged with myc Lane 2 : Transfected CHO cells expressing human Pick1 tagged with myc
Lysates/proteins at 20 µg per lane.
Secondary Lane 2 : An HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution
IHC image of ab3420 staining in rat adult brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3420, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunocytochemistry/ Immunofluorescence - PICK1 antibody (ab3420)This image is courtesy of an Anonymous Abreview.
ab3420 staining PICK1 in CHO cells stably transfected with Human PICK1 by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde and permeabilized in 0.5% Triton X-100 prior to blocking in 10% serum for 1 hour at room temperature. The primary antibody was diluted 1/1000 in PBS, 0.5% Triton, 10% FCS and incubated with the sample for 2 hours at °C. The secondary antibody was Alexa Fluor® 488-conjugated Goat anti-Rabbit polyclonal, diluted 1/500.
Western blot of PICK1 on AtT20 cell lysate using ab3420.
References for Anti-PICK1 antibody (ab3420)
This product has been referenced in:
Ma TM et al. Pathogenic disruption of DISC1-serine racemase binding elicits schizophrenia-like behavior via D-serine depletion. Mol Psychiatry : (2012).
Read more (PubMed: 22801410) »
Thorsen TS et al. Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD. Proc Natl Acad Sci U S A107:413-8 (2010).
Read more (PubMed: 20018661) »