Overview

  • Product name

    Anti-NCAM1 antibody [EPR21827]
    See all NCAM1 primary antibodies
  • Description

    Rabbit monoclonal [EPR21827] to NCAM1
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Mouse NCAM1 aa 1-350. The exact sequence is proprietary.
    Database link: P13595

  • Positive control

    • WB: Neuro-2a whole cell lysate; Mouse and rat brain and cerebellum lysates. IHC-P: Mouse stomach and cerebrum tissues; Rat colon tissue. ICC/IF: Neuro-2a cells. Flow Cyt: Neuro-2a cells. IP: Mouse and rat brain lysates.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab220360 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
WB 1/1000. Detects a band of approximately 120-200 kDa (predicted molecular weight: 95 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
IP 1/30.
Flow Cyt 1/600.

Target

  • Function

    This protein is a cell adhesion molecule involved in neuron-neuron adhesion, neurite fasciculation, outgrowth of neurites, etc.
  • Sequence similarities

    Contains 2 fibronectin type-III domains.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • antigen MSK39 identified by monoclonal antibody 5.1H11 antibody
    • antigen recognized by monoclonal antibody 5.1H11 antibody
    • CD56 antibody
    • cell adhesion molecule, neural, 1 antibody
    • MSK 39 antibody
    • MSK39 antibody
    • N-CAM-1 antibody
    • NCAM 1 antibody
    • NCAM antibody
    • NCAM C antibody
    • NCAM-1 antibody
    • NCAM1 antibody
    • NCAM1_HUMAN antibody
    • NCAMC antibody
    • Neural cell adhesion molecule 1 antibody
    • Neural cell adhesion molecule NCAM antibody
    • OTTHUMP00000235666 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling NCAM1 with ab220360 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Positive staining of ganglion (arrow) in mouse stomach (PMID: 1705171). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rab it IgG H&L (HRP), Ready to use.

  • All lanes : Anti-NCAM1 antibody [EPR21827] (ab220360) at 1/1000 dilution

    Lane 1 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate
    Lane 2 : L-929 (mouse connective tissue fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 95 kDa
    Observed band size: 140 kDa
    why is the actual band size different from the predicted?


    Exposure time: 70 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Negative control: L-929 (PMID: 9696812)

  • All lanes : Anti-NCAM1 antibody [EPR21827] (ab220360) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse cerebellum lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Rat cerebellum lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 95 kDa
    Observed band size: 120,140,180 kDa why is the actual band size different from the predicted?



    Exposure time: Lanes 1-2: 26 seconds; Lane 3: 70 seconds; Lane 4: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The 120,140 and 180kDa bands are different isoforms as reported in the literature (PMID:26288071).

  • Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling NCAM1 with ab220360 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Positive staining on mouse cerebrum (PMID: 1705171). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rab it IgG H&L (HRP), Ready to use.

  • Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling NCAM1 with ab220360 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Positive staining of ganglia (arrows) in rat colon (PMID: 1705171). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rab it IgG H&L (HRP), Ready to use.

  • Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling NCAM1 with ab220360 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Neuro-2a cell line.

    Negative control: L-929 PMID: 9696812).

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Flow cytometric analysis of L-929 (mouse connective tissue fibroblast cell line) cell line (left panel) and Neuro-2a (mouse neuroblastoma cell line) cell line (right panel) labeling NCAM1 with ab220360 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. 

    Gated on viable cells. Negative control: L-929.

  • NCAM1 was immunoprecipitated from 0.35 mg of mouse brain lysate with ab220360 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220360 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Mouse brain lysate 10 µg (Input). 

    Lane 2: ab220360 IP in mouse brain lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220360 in mouse brain lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

    The 120,140 and 180kDa bands are different isoforms as reported in the literature (PMID: 26288071).

  • NCAM1 was immunoprecipitated from 0.35 mg of rat brain lysate with ab220360 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220360 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Rat brain lysate 10 µg (Input). 

    Lane 2: ab220360 IP in rat brain lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220360 in rat brain lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

    The 120,140 and 180kDa bands are different isoforms as reported in the literature (PMID: 26288071).

References

ab220360 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab220360.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up