Overview

  • Product name

    Anti-NCAM1 antibody [CAL53]
    See all NCAM1 primary antibodies
  • Description

    Rabbit monoclonal [CAL53] to NCAM1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human NCAM1 aa 740-858. The exact sequence is proprietary.
    Database link: P13591

  • Positive control

    • WB: Human brain and cerebellum lysates; mouse cerebellum lysate; rat brain lysate; Neuro-2a whole cell lysate. IHC-P: Human cerebrum, glioma, pancreas and pancreatic carcinoma tissue; mouse cerebrum tissue; rat cerebrum tissue. ICC/IF: Neuro-2a cells. IP: SH-SY5Y whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab237708 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 140-180 kDa (predicted molecular weight: 94 kDa).
ICC/IF 1/50.
IP 1/20.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    Images

    • Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human glioma is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

       

    • Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human cerebrum is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

       

    • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on islet in the human pancreas is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

       

    • Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on islet in the mouse cerebrum is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

       

    • Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on islet in the rat cerebrum is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling NCAM1 with ab237708 at 1/50 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 cells. 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).

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

      Negative control: L-929 (PMID:9696812).

    • NCAM1 was immunoprecipitated from 0.35 mg of SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate with ab237708 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab237708 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

      Lane 1: SH-SY5Y whole cell lysate 10 μg (Input).
      Lane 2: ab237708 IP in SH-SY5Y whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237708 in SH-SY5Y whole cell lysate.

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

    • All lanes : Anti-NCAM1 antibody [CAL53] (ab237708) at 1/1000 dilution

      Lane 1 : Human brain lysate
      Lane 2 : Human cerebellum lysate
      Lane 3 : Neuro-2a (mouse neuroblastoma cell line0 whole cell lysate
      Lane 4 : Mouse cerebellum lysate
      Lane 5 : Rat brain lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 94 kDa
      Observed band size: 140-180 kDa why is the actual band size different from the predicted?


      Exposure time: 26 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Formalin-fixed, paraffin-embedded human pancreatic carcinoma tissue stained for NCAM1 using ab237708 at 0.25 µg/ml in immunohistochemical analysis.

    References

    ab237708 has not yet been referenced specifically in any publications.

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