Overview

  • Product name
    Anti-Mannosidase II antibody
    See all Mannosidase II primary antibodies
  • Description
    Rabbit polyclonal to Mannosidase II
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IFmore details
    Unsuitable for: IP or WB
  • Species reactivity
    Reacts with: Human, Monkey
    Does not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide to internal sequence (Human) conjugated to KLH.

  • Positive control
    • ICC: COS7 cells

Properties

Applications

Our Abpromise guarantee covers the use of ab12277 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
ICC/IF 1/200.

Use with methanol fixed cells. 4% paraformaldehyde fixation reduces signal.

  • Application notes
    Is unsuitable for IP or WB.
  • Target

    • Function
      Catalyzes the first committed step in the biosynthesis of complex N-glycans. It controls conversion of high mannose to complex N-glycans; the final hydrolytic step in the N-glycan maturation pathway.
    • Pathway
      Protein modification; protein glycosylation.
    • Sequence similarities
      Belongs to the glycosyl hydrolase 38 family.
    • Cellular localization
      Golgi apparatus membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • 3-1 antibody
      • 6-alpha-mannosidase antibody
      • Alpha mannosidase 2 antibody
      • alpha Mannosidase II antibody
      • Alpha-mannosidase 2 antibody
      • AMAN II antibody
      • Golgi alpha mannosidase II antibody
      • Golgi alpha-mannosidase II antibody
      • Golgi integral membrane protein 7 antibody
      • GOLIM7 antibody
      • MA2A1_HUMAN antibody
      • MAN II antibody
      • Man2a1 antibody
      • MANA 2 antibody
      • MANA2 antibody
      • MANII antibody
      • Mann II antibody
      • Mannosidase alpha class 2A member 1 antibody
      • Mannosidase Two antibody
      • Mannosidase, alpha type II antibody
      • Mannosidase, alpha, II antibody
      • Mannosyl oligosaccharide 1 3 1 6 alpha mannosidase antibody
      • Mannosyl oligosaccharide 1,3 1,6 alpha mannosidase antibody
      • Mannosyl-oligosaccharide 1 antibody
      see all

    Images

    • COS7 cells were fixed with cold methanol for 10 minutes followed by incubation with ab12277 (1/200) in 5% normal goat serum. FITC-conjugated goat anti-rabbit was used to visualise bound rabbit antibodies. Nuclei were stained with DAPI.
    • ICC/IF image of ab12277 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab12277, 1/1000 dilution) 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.

    References

    This product has been referenced in:
    • Jiang P  et al. The Protein Encoded by the CCDC170 Breast Cancer Gene Functions to Organize the Golgi-Microtubule Network. EBioMedicine 22:28-43 (2017). Read more (PubMed: 28687497) »
    • Vashi N  et al. Ca2+-dependent Focal Exocytosis of Golgi-derived Vesicles Helps Phagocytic Uptake in Macrophages. J Biol Chem 292:5144-5165 (2017). Read more (PubMed: 28174296) »
    See all 24 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Rat hepatocytes)
    Permeabilization
    Yes - 0.2% Triton X-100
    Specification
    Rat hepatocytes
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Formaldehyde

    Dr. Armen Petrosyan

    Verified customer

    Submitted Aug 24 2015

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HepG2 cells)
    Permeabilization
    Yes - 0.2% Triton X-100
    Specification
    HepG2 cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Formaldehyde

    Dr. Armen Petrosyan

    Verified customer

    Submitted Aug 24 2015

    Answer

    Vielen Dank für Ihre Anfrage.
    Leider ist es nicht möglich die Konzentration von ab12277 und ab136497 zu bestimmen.
    Bei ab12277 und ab136497 handelt es sich um Vollserum.
    Die Konzentration von Antikörpern wird üblicher Weise über die Proteinmenge bestimmt. Vollserum enthält aber nicht nur den spezifischen Antikörper sondern auch andere Proteine (und Antikörper) und macht deshalb eine Konzentrationsbestimmung unmöglich.
    Wir empfehlen als Anhaltspunkt eine Konzentration von 1mg/ml anzunehmen für den spezifischen Antikörper im Vollserum.
    Es tut mir leid, dass ich Ihnen keine genaueren Angaben machen konnte und hoffe diese Information ist dennoch hilfreich.

    Read More

    Answer

    Thank you for your questions. I apologize for the delay in replying but I was unable to contact the laboratory that produces this antibody for information about FACS testing, or for more information about the immunogen.

    I expect that the antibody will be suitable for FACS, given that it has been used to stain cells for microscopy. The plasma membrane of the cells will need to be permeabilized with methanol as described in the caption of the photo of ICC/IF on the datasheet, to allow the antibody access to the mannosidase in the ER.

    It is very likely that the immunogen for the antibody was derived from the lumenal region. According to the description of the human protein at the UniProt database, the cytoplasmic portion is amino acids 1-5, the transmembrane region is 5-26, and the lumenal portion is 27-1144.

    http://www.uniprot.org/uniprot/Q16706

    Read More

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

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