Overview

  • Product name
    Anti-Syntaxin 6 antibody [3D10]
    See all Syntaxin 6 primary antibodies
  • Description
    Mouse monoclonal [3D10] to Syntaxin 6
  • Tested applications
    Suitable for: Flow Cyt, WB, IP, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Human
  • Immunogen

    Recombinant rat syntaxin 6 fusion protein.

  • Epitope
    The epitope for this antibody has been localized to the amino terminus (residues 1-25) of Syntaxin 6.
  • Positive control
    • Mouse Brain Tissue Extract, rat Brain Tissue Extract.
  • 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. We would recommend antibody ab140607 as a replacement.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.09% Sodium Azide
    Constituents: 50% Glycerol, PBS, pH 7.2
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Monoclonal
  • Clone number
    3D10
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab12370 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. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 30.6 kDa).
IP Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.

Target

Images

  • ab12370 at 1/200 staining a mouse macrophage cell line by ICC/IF. The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 24 hours. A FITC conjugated donkey anti-mouse antibody was used as the secondary.

    See Abreview

  • All lanes : Anti-Syntaxin 6 antibody [3D10] (ab12370) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Brain (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 30.6 kDa
    Observed band size : 30.6 kDa
    Additional bands at : 26 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes


  • Predicted band size : 30.6 kDa

    Image from Jonas MC et al, J Cell Sci. 2010 Oct 1;123(Pt 19):3378-88. Epub 2010 Sep 7, Fig 1, doi:10.1242/jcs.068841.

  • Syntaxin 6 was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 10µg of Mouse monoclonal to Syntaxin 6 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab12370.
    Secondary: Protein G-HRP at 1/500 dilution.
    Band: 25kDa:Syntaxin 6.
  • Overlay histogram showing PC12 cells stained with ab12370 (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 (ab12370, 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.

References

This product has been referenced in:
  • Sotthibundhu A  et al. Rapamycin regulates autophagy and cell adhesion in induced pluripotent stem cells. Stem Cell Res Ther 7:166 (2016). ICC/IF ; Human . Read more (PubMed: 27846905) »
  • Hao Z  et al. Impaired maturation of large dense-core vesicles in muted-deficient adrenal chromaffin cells. J Cell Sci 128:1365-74 (2015). Read more (PubMed: 25673877) »

See all 15 Publications for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (10 %)
Sample
Guinea pig Tissue lysate - whole (Liver)
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Mrs. Monika Schumacher

Verified customer

Submitted Jan 23 2015

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (10 %)
Sample
Human Cell lysate - whole cell (Several cell lines)
Specification
Several cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Mrs. Monika Schumacher

Verified customer

Submitted Jan 16 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse macrophage cell line)
Specification
mouse macrophage cell line
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1.5%
Username

Abcam user community

Verified customer

Submitted Feb 27 2007

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

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