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Anti-Kidins220 Antibody

Kinase D-interacting substrate of 220 kDa, Ankyrin repeat-rich membrane-spanning protein, ARMS
Cat #: APZ-017
Alternative Name Kinase D-interacting substrate of 220 kDa, Ankyrin repeat-rich membrane-spanning protein, ARMS
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)PRHSNLSWQSQTRR, corresponding to amino acid residues 1335-1348 of rat Kidins220 (Accession Q9EQG6). Intracellular, C-terminus.
Accession (Uniprot) Number Q9EQG6
Gene ID 116478
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse, human – identical.
RRID AB_2756775.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ih, wb
May also work in: ic*, ifc*, ip*
Western blot
  • Western blot analysis of mouse brain membranes (lanes 1 and 3) and rat brain synaptosomal fraction (lanes 2 and 4): 
    Western blot analysis of mouse brain membranes (lanes 1 and 3) and rat brain synaptosomal fraction (lanes 2 and 4): 
    1,2. Anti-Kidins220 Antibody (#APZ-017), (1:200).
    3,4. Anti-Kidins220 Antibody, preincubated with Kidins220 Blocking Peptide (#BLP-PZ017).
Immunohistochemistry
  • Expression of Kidins220 in mouse hippocampus and cerebellum
    Expression of Kidins220 in mouse hippocampus and cerebellum
    Immunohistochemical staining of perfusion-fixed frozen mouse brain sections with Anti-Kidins220 Antibody (#APZ-017), (1:1000), followed by goat-anti-rabbit-AlexaFluor-488. A. Kidins220 staining (green) in hippocampal sections is seen in neurons in the pyramidal layer (Pyr) and stratum oriens (OR) (arrows). B. Staining in cerebellum sections shows labeling (green) in neurons in the Purkinje cell layer (horizontal arrows). Cell nuclei are stained with DAPI (blue).
References
  1. Neubrand, V.E. et al. (2012) J. Cell Sci. 125, 1845.
  2. Arevalo, J.C. et al. (2004) EMBO J. 23, 2358.
Scientific background

The Kidins220 (Kinase D-interacting substrate of 220 kDa) or ARMS (Ankyrin repeat-rich membrane-spanning) protein was originally identified as a substrate for protein kinase D (PKD) in neural cells. Kidins220 is a member of the Kidins220/ARMS and PifA (KAP) family of P-loop nucleotide phosphatases (NTPases).

Kidins220 is an integral membrane protein, containing four transmembrane segments in the central part of the molecule, and N- and C-terminal tails both exposed to the cytoplasm. Kidins220 possesses numerous domains that mediate protein–protein interactions. The N-terminus of Kidins220 contains eleven ankyrin repeats which form a concave binding surface that is accessible to molecular partners. Kidins220 contains Walker A and Walker B (involved in the binding of nucleotides) motifs in its juxta-membrane regions flanking the N- and C-termini. The C-terminus of Kidins220 includes several protein–protein interaction domains that are involved in inter- and intra-molecular interactions. The last four amino acids of Kidins220 constitute a PDZ-binding motif that binds the PDZ domain of α-syntrophin, thereby forming a complex that is associated with the EphA4 receptor at the neuromuscular junction1.

Neurotrophin signaling is responsible for a wide variety of neuronal functions and is dependent upon two different transmembrane receptor types; the Trk receptor tyrosine kinase and the p75 neurotrophin receptor that belongs to the TNF receptor family. Kidins220 interacts with TrkATrkBTrkC receptors, and with p75NTR. Kidinis220 is rapidly tyrosine phosphorylated after binding of neurotrophins to Trk receptors and provides a docking site for the CrkL–C3G complex, resulting in Rap1-dependent sustained ERK activation2.

Kidins220 also associates with NMDA receptors. Upon overstimulation of the NMDA receptor, activity-dependent Ca2+ influx through NMDA receptors leads to a decrease in Kidins220 levels owing to transcriptional downregulation and protein cleavage by calpain1.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 23/08/2020

Anti-Kidins220 Antibody (#APZ-017) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize Kidins220 from human, mouse, and rat samples.

For research purposes only, not for human use
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