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RecombinantRabMAb

Recombinant Anti-NCAM1 antibody [EPR21827] (ab220360)

Reviews (2) Submit a question References (8)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Western blot - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Western blot - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Flow Cytometry - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] (ab220360)
  • Anti-NCAM1 antibody [EPR21827] (ab220360)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21827] to NCAM1
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 APC Carrier Free FITC PE

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. This information is proprietary to Abcam and/or its suppliers.

  • 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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

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)
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)
1/600.
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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)

      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.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Western blot - Anti-NCAM1 antibody [EPR21827] (ab220360)
      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)

    • Western blot - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Western blot - Anti-NCAM1 antibody [EPR21827] (ab220360)
      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).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)

      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.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] (ab220360)

      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.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR21827] (ab220360)

      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 Cytometry - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Flow Cytometry - Anti-NCAM1 antibody [EPR21827] (ab220360)

      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.

    • Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] (ab220360)

      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).

    • Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] (ab220360)
      Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] (ab220360)

      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).

    • Anti-NCAM1 antibody [EPR21827] (ab220360)
      Anti-NCAM1 antibody [EPR21827] (ab220360)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (8)

    Publishing research using ab220360? Please let us know so that we can cite the reference in this datasheet.

    ab220360 has been referenced in 8 publications.

    • Hirata T  et al. Loss of the N-acetylgalactosamine side chain of the GPI-anchor impairs bone formation and brain functions and accelerates the prion disease pathology. J Biol Chem 298:101720 (2022). PubMed: 35151686
    • Song G  et al. Single-cell transcriptomic analysis suggests two molecularly subtypes of intrahepatic cholangiocarcinoma. Nat Commun 13:1642 (2022). PubMed: 35347134
    • Liu T  et al. SPIONs mediated magnetic actuation promotes nerve regeneration by inducing and maintaining repair-supportive phenotypes in Schwann cells. J Nanobiotechnology 20:159 (2022). PubMed: 35351151
    • Yu J  et al. Effects and potential mechanism of Ca2+/calmodulin‑dependent protein kinase II pathway inhibitor KN93 on the development of ovarian follicle. Int J Mol Med 50:N/A (2022). PubMed: 35929517
    • Duregotti E  et al. Reduced secretion of neuronal growth regulator 1 contributes to impaired adipose-neuronal crosstalk in obesity. Nat Commun 13:7269 (2022). PubMed: 36433953
    • Sun JH  et al. Nerve bundle formation during the promotion of peripheral nerve regeneration: collagen VI-neural cell adhesion molecule 1 interaction. Neural Regen Res 17:1023-1033 (2022). PubMed: 34558529
    • Woo J  et al. High-throughput and high-efficiency sample preparation for single-cell proteomics using a nested nanowell chip. Nat Commun 12:6246 (2021). PubMed: 34716329
    • Wang X  et al. The intact parasympathetic nerve promotes submandibular gland regeneration through ductal cell proliferation. Cell Prolif 54:e13078 (2021). PubMed: 34101282

    Customer reviews and Q&As

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    1-2 of 2 Abreviews

    Flow Cytometry abreview for Anti-NCAM1 antibody [EPR21827]

     Below Average
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Cow Cell (MSCs bovine cells)
    Permeabilization
    No
    Gating Strategy
    All cells
    Specification
    MSCs bovine cells
    Preparation
    Cell harvesting/tissue preparation method: 100ul of blood was added to each well (96 well) and the first antibody was added, after incubation and washes the secondary antibody was added and incubated (in the dark) following washes. Afterward, the sample was read by the flow cytometer
    Sample buffer: 3% BSA in PBS
    Fixation
    The cells weren't fixated
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Sep 30 2022

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-NCAM1 antibody [EPR21827]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Colon)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH9
    Permeabilization
    No
    Specification
    Colon
    Blocking step
    (agent) for 18 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
    Fixative
    10% NBF
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Vicki Shone

    Verified customer

    Submitted Dec 16 2020

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