Overview

  • Product name
  • Description
    Mouse monoclonal to MGAT5
  • Tested applications
    Suitable for: WB, ELISA, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Hamster
  • Immunogen

    Recombinant fragment: LAEPGQSCKQ VCQESQLICE PSFFQHLNKD KDMLKYKVTC QSSELAKDIL VPSFDPKNK HCVFQGDLLL FSCAGAHPRH QRVCPCRDFI KGQVALCKD, corresponding to amino acids 642 - 739 of Human MGAT5 (NP_002401) with a proprietary tag

  • Positive control
    • Jurkat cell lysate
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause.

Properties

Applications

Our Abpromise guarantee covers the use of ab87977 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 Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 85 kDa.
ELISA Use at an assay dependent concentration.
Flow Cyt Use 0.1-1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Catalyzes the addition of N-acetylglucosamine in beta 1-6 linkage to the alpha-linked mannose of biantennary N-linked oligosaccharides. It is one of the most important enzymes involved in the regulation of the biosynthesis of glycoprotein oligosaccharides.
  • Pathway
    Protein modification; protein glycosylation.
  • Sequence similarities
    Belongs to the glycosyltransferase 18 family.
  • Cellular localization
    Golgi apparatus membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A antibody
    • 6-N-acetylglucosaminyltransferase antibody
    • Alpha 1,3(6) mannosylglycoprotein antibody
    • Alpha 1,6 mannosylglycoprotein 6 beta N acetylglucosaminyltransferase antibody
    • Alpha mannoside beta 1,6 N acetylglucosaminyltransferase antibody
    • Alpha-1 antibody
    • Alpha-mannoside beta-1 antibody
    • Beta 1,6 N acetyl glucosaminyltransferase antibody
    • GGNT5 antibody
    • GlcNAc T V antibody
    • GlcNAc-T V antibody
    • GNT V antibody
    • GNT VA antibody
    • GNT-V antibody
    • GNTV antibody
    • GNTVA antibody
    • Mannoside acetylglucosaminyltransferase 5 antibody
    • Mannosyl (alpha 1,6) glycoprotein beta 1,6 N acetyl glucosaminyltransferase antibody
    • Mgat5 antibody
    • MGT5A_HUMAN antibody
    • N acetylglucosaminyl transferase V antibody
    • N acetylglucosaminyltransferase V mannosyl (alpha 1,6) glycoprotein antibody
    • N-acetylglucosaminyl-transferase V antibody
    see all

Images

  • Anti-MGAT5 antibody (ab87977) at 1/2000 dilution + Human RWPE prostate cells at 20 µg

    Secondary
    Donkey anti-mouse polyclonal HRP conjugate at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 85 kDa
    Additional bands at : 55 kDa (possible non-specific binding),95 kDa (possible post-translational modification).

    Exposure time : 40 seconds

    Image is courtesy of an AbReview submitted by Armen Petrosyan.

    Blocking was via 5% milk in PBST incubated for 1 hour at 22°C

    See Abreview

  • Overlay histogram showing Jurkat cells stained with ab87977 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab87977, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • Anti-MGAT5 antibody (ab87977) at 1 µg/ml + recombinant protein fragment with a proprietary tag (the immunogen) at 0.1 µg

    Secondary
    Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/5000 dilution

    Predicted band size : 85 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
    The recombinant protein used as the immunogen has a predicted size of 37 kDa.
  • Anti-MGAT5 antibody (ab87977) at 1 µg/ml + jurkat cell lysate at 25 µg

    Secondary
    Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

    Predicted band size : 85 kDa

References

ab87977 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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
Username

Dr. Armen Petrosyan

Verified customer

Submitted Nov 27 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (RWPE - prostate cells)
Gel Running Conditions
Reduced Denaturing (8% SDS-PAGE)
Loading amount
20 µg
Specification
RWPE - prostate cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Armen Petrosyan

Verified customer

Submitted Oct 09 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (PC-3 prostate cancer cells)
Permeabilization
Yes - 0.2% Triton X-100
Specification
PC-3 prostate cancer cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

Submitted Aug 18 2015

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

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