Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker

• Flow Cyt

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Flow Cytometry - Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker (AB12370)

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.

This image was generated using the ascites version of the product.

• IP

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Immunoprecipitation - Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker (AB12370)

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 70^oC; 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.

This image was generated using the ascites version of the product.

All lanes:

Immunoprecipitation - Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker (ab12370)

Predicted band size: 29 kDa

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• WB

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Western blot - Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker (AB12370)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker (ab12370) at 1 µg/mL

Lane 1:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 2:

Brain (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 29 kDa

Observed band size: 26 kDa,30.6 kDa

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Exposure time: 8min

• WB

PubMed

Western blot - Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker (AB12370)

Intracellular membranes from CHO cells were separated on a 10–24% discontinuous Nycodenz gradient. Western Blotting was then performed using a range of antibodies including ab12370 at a 1/1000 dilution. An HRP-conjugated anti-mouse secondary was used at a 1/6000 dilution.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker (ab12370)

Predicted band size: 29 kDa

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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.