The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 mg/ml. Detects a band of approximately 152 kDa (predicted molecular weight: 148 kDa).
Use at an assay dependent concentration.
May play a role in the formation of functional distinct domains critical for saltatory conduction of nerve impulses in myelinated nerve fibers. Seems to demarcate the juxtaparanodal region of the axo-glial junction.
Predominantly expressed in nervous system.
Involvement in disease
Defects in CNTNAP2 are the cause of cortical dysplasia-focal epilepsy syndrome (CDFES) [MIM:610042]. Affected individuals manifest cortical dysplasia, focal epilepsy, relative macrocephaly, and diminished deep-tendon reflexes. Intractable focal seizures begin in early childhood, after which language regression, hyperactivity, impulsive and aggressive behavior, and mental retardation develop. Genetic variations in CNTNAP2 influences susceptibility to autism type 15 (AUTS15) [MIM:612100]. Autism is a neurodevelopmental disorder characterized by disturbance in language, perception and socialization. The disorder is classically defined by a triad of limited or absent verbal communication, a lack of reciprocal social interaction or responsiveness, and restricted, stereotypical, and ritualized patterns of interests and behavior. Note=A chromosomal aberration involving CNTNAP2 is found in a patient with autism spectrum disorder. Paracentric inversion 46,XY,inv(7)(q11.22;q35). The inversion breakpoints disrupt the genes AUTS2 and CNTNAP2.
Belongs to the neurexin family. Contains 2 EGF-like domains. Contains 1 F5/8 type C domain. Contains 1 fibrinogen C-terminal domain. Contains 4 laminin G-like domains.
All lanes : Anti-Caspr2 antibody (ab33994) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate at 20 µg Lane 2 : Brain (Rat) Tissue Lysate at 20 µg Lane 3 : Brain (Human) Tissue Lysate at 20 µg Lane 4 : Sciatic Nerve (Mouse) Lysate at 7.5 µg Lane 5 : Sciatic Nerve (Rat) Lysate at 20 µg
Secondary All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Caspr2 was immunoprecipitated using 0.5 mg mouse brain whole tissue lysate, 5µg of Rabbit polyclonal to Caspr2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10 minutes, mouse brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 minutes under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10 minutes at 70°C; 10 µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab33994. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Band: 152 kDa: Caspr2.
ICC/IF image of ab33994 stained SHSY5Y cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab33994, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Dawes JM et al. Immune or Genetic-Mediated Disruption of CASPR2 Causes Pain Hypersensitivity Due to Enhanced Primary Afferent Excitability. Neuron97:806-822.e10 (2018).
Read more (PubMed: 29429934) »
Saifetiarova J et al. Early and Late Loss of the Cytoskeletal Scaffolding Protein, Ankyrin G Reveals Its Role in Maturation and Maintenance of Nodes of Ranvier in Myelinated Axons. J Neurosci37:2524-2538 (2017).
Read more (PubMed: 28148727) »