Overview

  • Product name

  • Description

    Rabbit polyclonal to NSMase2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Rat
    Predicted to work with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human NSMase2 aa 600 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab95417)

  • Positive control

    • This antibody gave a positive signal in Rat Brain tissue lysate and human lung tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab85017 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 a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 71 kDa (predicted molecular weight: 71 kDa).

Target

  • Function

    Catalyzes the hydrolysis of sphingomyelin to form ceramide and phosphocholine. Ceramide mediates numerous cellular functions, such as apoptosis and growth arrest, and is capable of regulating these 2 cellular events independently. Also hydrolyzes sphingosylphosphocholine. Regulates the cell cycle by acting as a growth suppressor in confluent cells. Probably acts as a regulator of postnatal development and participates in bone and dentin mineralization.
  • Tissue specificity

    Predominantly expressed in brain.
  • Sequence similarities

    Belongs to the neutral sphingomyelinase family.
  • Developmental stage

    Up-regulated during G0/G1 phases.
  • Cellular localization

    Golgi apparatus membrane. Cell membrane. May localize to detergent-resistant subdomains of Golgi membranes of hypothalamic neurosecretory neurons. According to PubMed:15051724, it localizes to plasma membrane in confluent contact-inhibited cells.
  • Information by UniProt
  • Database links

  • Alternative names

    • Cca1 antibody
    • Confluent 3Y1 cell-associated protein 1 antibody
    • Neutral sphingomyelinase 2 antibody
    • Neutral sphingomyelinase II antibody
    • NSMA2_HUMAN antibody
    • nSMase-2 antibody
    • nSMase2 antibody
    • SMPD3 antibody
    • Sphingomyelin phosphodiesterase 3 antibody
    • sphingomyelin phosphodiesterase 3, neutral membrane (neutral sphingomyelinase II) antibody
    see all

Images

  • Anti-NSMase2 antibody (ab85017) at 1 µg/ml + Brain (Rat) Tissue Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 71 kDa
    Observed band size: 71 kDa
    Additional bands at: 102 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes
  • ICC/IF image of ab85017 stained PC12 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85017, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) PC12 cells at 5µg/ml.
  • IHC image of ab85017 staining in human lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85017, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:

See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Dorsal skin)
Permeabilization
Yes - 0.25% Triton
Specification
Dorsal skin
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1%
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 08 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (brain)
Specification
brain
Blocking step
BSA/NGS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Fixative
Paraformaldehyde

Matthieu Lecuyer

Verified customer

Submitted Jul 30 2018

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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