Overview

  • Product name

  • Description

    Rabbit polyclonal to Caspr
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, IP, IHC-FoFrmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse Caspr aa 1350 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab34150)

  • Positive control

    • Rat brain whole cell lysate and PC12 cytoplasmic lysate. ICC/IF:SKNSH cell line

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS. pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab34151 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
WB 1/250. Detects a band of approximately 180 kDa (predicted molecular weight: 156 kDa).Can be blocked with Mouse Caspr peptide (ab34150).
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration.
IHC-FoFr 1/3000.

Target

  • Function

    Seems to 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 paranodal region of the axo-glial junction. In association with contactin may have a role in the signaling between axons and myelinating glial cells.
  • Tissue specificity

    Predominantly expressed in brain. Weak expression detected in ovary, pancreas, colon, lung, heart, intestine and testis.
  • Sequence similarities

    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.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Caspr antibody
    • Caspr1 antibody
    • CNTNAP antibody
    • Cntnap1 antibody
    • CNTP1_HUMAN antibody
    • Contactin associated protein 1 antibody
    • Contactin-associated protein 1 antibody
    • MHDNIV antibody
    • NCP1 antibody
    • Neurexin 4 antibody
    • Neurexin IV antibody
    • Neurexin-4 antibody
    • Nrxn4 antibody
    • p190 antibody
    • Paranodin antibody
    see all

Images

  • ab34151 stained in SKNSH cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab34151 at 1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • All lanes : Anti-Caspr antibody (ab34151) at 1/250 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : CNTNAP1 (Caspr) knockout HAP1 whole cell lysate
    Lane 3 : MEF whole cell lysate
    Lane 4 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 156 kDa
    Observed band size: 180 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab34151 observed at 180 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab34151 was shown to recognize Caspr in wild-type HAP1 cells as signal was lost at the expected MW in CNTNAP1 (Caspr) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CNTNAP1 (Caspr) knockout samples were subjected to SDS-PAGE. ab34151 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/250 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Anti-Caspr antibody (ab34151) at 1/250 dilution + Brain (Rat) Whole Cell Lysate - normal tissue at 10 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 156 kDa
    Observed band size: 180 kDa why is the actual band size different from the predicted?
    Additional bands at: 58 kDa. We are unsure as to the identity of these extra bands.



    Caspr contains a number of potential glycosylation sites so it is thought that this is the reason it runs at 180kDa.
  • IHC-FoFR image of ab34151 stained sections of mouse brain (30 um). The tissues were from perfused fixed animals perfused with 4% PFA and postfixed 2h in the same fixative. They were cryoprotected in 30% sucrose and cut using a cryostat.

    See Abreview

  • Caspr was immunoprecipitated using 0.5mg Rat Brain whole tissue lysate, 5µg of Rabbit polyclonal to Caspr 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 10min, Rat Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab34151.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 180kDa: Caspr.
  • ab34151 staining Caspr in murine brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde. Samples were blocked with 20% serum for 1 hour at 20°C followed by incubation with the primary antibody at a 1/300 dilution for 12 hours at 20°C. An HRP-conjugated goat anti-rabbit polyclonal was used as the secondary antibody at a 1/200 dilution.

    See Abreview

References

This product has been referenced in:

  • Zhang SH  et al. A CASPR1-ATP1B3 protein interaction modulates plasma membrane localization of Na+/K+-ATPase in brain microvascular endothelial cells. J Biol Chem 294:6375-6386 (2019). Read more (PubMed: 30792309) »
  • Swire M & Ffrench-Constant C Oligodendrocyte-Neuron Myelinating Coculture. Methods Mol Biol 1936:111-128 (2019). Read more (PubMed: 30820896) »
See all 38 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Common marmoset Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Oct 28 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 14 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 13 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Postmortem brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Postmortem brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 13 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 20°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (brain)
Specification
brain
Permeabilization
No
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 21 2011

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Dorsal root ganglion)
Specification
Dorsal root ganglion

Dr. Sophie Pezet

Verified customer

Submitted Aug 03 2010

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