Overview

  • Product name
    Anti-NCAM1 antibody [RNL-1]
    See all NCAM1 primary antibodies
  • Description
    Mouse monoclonal [RNL-1] to NCAM1
  • Host species
    Mouse
  • Specificity
    Neural cell adhesion molecule (NCAM). ab9018 reacts with three isoforms of NCAM: NCAM-120, NCAM-140, and NCAM-180.
  • Tested applications
    Suitable for: Flow Cyt, IHC-FoFr, ICC/IF, ICC, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    This antibody was raised against the small lung cancer cell line NCI-H82.

  • Positive control
    • IHC-P: Mouse brain tissue. IHC-FoFr: Rat brain tissue. ICC/IF: Adult-derived human liver stem cells and hepatic stellate cells. Flow cytometry: HT1080 cells.
  • General notes
    Punctuate cytoplasmic staining is observed with ab9018. Some staining of red blood cells observed.

Properties

Applications

Our Abpromise guarantee covers the use of ab9018 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
Flow Cyt Use 2µg for 106 cells.

Paraformaldehyde or methanol fixed cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-FoFr 1/100 - 1/200. see detailed protocol
ICC/IF Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
WB 1/100 - 1/1000.
IHC-P Use at an assay dependent concentration.

Target

Images

  • Immunohistochemistry of H82 cells, showing staining for NCAM1 using ab9018 (1/500).

  • ab9018 staining NCAM in Mouse brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C; antigen retrieval was not performed. Samples were incubated with primary antibody (1/100) for 12 hours at 4°C. A Biotin-conjugated Rabbit anti-mouse polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • Immunocytochemical analysis of Adult-Derived Human Liver Stem Cells (ADHLSC) in the left image and Hepatic Stellate Cells (HSC) in the right image. ab9018 was used to label NCAM, which is expressed in the HSC. Cells were fixed in 3.5% paraformaldehyde for 15 minutes at room temperature. Blocking was with 3.3% hydrogen peroxide for 3 minutes. Permeabilization with D-PBS and 1% Triton X-100 for 10 minutes. Cells incubated with ab9018 at 1/100 for 1 hour. DAB staining. Counterstaining with Mayer's hematoxylin. 

  • Overlay histogram showing HT1080 cells stained with ab9018 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9018, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HT1080 cells fixed with methanol (5 min) used under the same conditions.

    Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • ab9018 at a dilution of 1/500, staining NCAM (Alexa 488 secondary at 1/2000) on rat brain tissue (30µm thick coronal sections) in free floating IHC (see protocol link for detailed description). Images showing neuron body and processes: [A] 40x objective and [B] punctate cytoplasmic staining; 20x objective and [C] neuron + processes; x20 objective. No labeling observed following omission of primary antibody. Images coloured in Photoshop.

    NB: We recommend indirect amplification of immunofluorescence for ab9018

    Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).

References

This product has been referenced in:
  • Feng Q  et al. Integrative proteomics and immunochemistry analysis of the factors in the necrosis and repair in acetaminophen-induced acute liver injury in mice. J Cell Physiol 234:6561-6581 (2019). Read more (PubMed: 30417486) »
  • Eguchi T  et al. Organoids with cancer stem cell-like properties secrete exosomes and HSP90 in a 3D nanoenvironment. PLoS One 13:e0191109 (2018). Read more (PubMed: 29415026) »
See all 18 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Sep 14 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cultured Cells (HT1080)
Specification
HT1080
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100

Abcam user community

Verified customer

Submitted Mar 17 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - other (stem cells, neural precursors)
Specification
stem cells, neural precursors
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jan 30 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (stem cells, glial precursors)
Specification
stem cells, glial precursors
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jan 30 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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