Anti-SNAP29 antibody [EPR9198(2)] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SNAP29 antibody [EPR9198(2)] - BSA and Azide free (AB240248)

Immunofluorescent analysis of -20? acetone fixed Hela cells staining SNAP29 using ab181151 at 1/250 dilution and Alexa Fluor®555 stained Goat anti Rabbit IgG at 1/250 as a secondary antibody (red). Dapi counterstain (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181151).

• WB

Lab

Western blot - Anti-SNAP29 antibody [EPR9198(2)] - BSA and Azide free (AB240248)

This data was developed using the same antibody clone in a different buffer formulation (ab181151).

Lanes 1-3 : Merged signal (red and green). Green - ab181151 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.

ab181151 Anti-SNAP29 antibody [EPR9198(2)] was shown to specifically react with SNAP29 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265289 (knockout cell lysate ab257693) was used. Wild-type and SNAP29 knockout samples were subjected to SDS-PAGE. ab181151 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-SNAP29 antibody [EPR9198(2)] (<a href='/en-us/products/primary-antibodies/snap29-antibody-epr91982-ab181151'>ab181151</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SNAP29 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SNAP29 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-snap29-knockout-hela-cell-line-ab265289'>ab265289</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 29 kDa

Observed band size: 29 kDa

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• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SNAP29 antibody [EPR9198(2)] - BSA and Azide free (AB240248)

Intracellular flow cytometric analysis of 2% paraformaldehyde fixed Jurkat cells staining SNAP29 using ab181151 at 1/10 dilution, and FITC conjugated Goat anti rabbit IgG at 1/150 as a secondary antibody (red curve). Rabbit monoclonal IgG was used as the negative control (green curve).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181151).

• IP

Supplier Data

Immunoprecipitation - Anti-SNAP29 antibody [EPR9198(2)] - BSA and Azide free (AB240248)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181151).

Western blot analysis of SNAP29 in immunoprecipitation pellets from Jurkat cell lysate, using ab181151 at a 1/50 dilution.

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as a secondary antibody at 1/1000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-SNAP29 antibody [EPR9198(2)] (<a href='/en-us/products/primary-antibodies/snap29-antibody-epr91982-ab181151'>ab181151</a>) at 1/50 dilution

Lane 1:

Precipitate from Jurkat lysate using <a href='/en-us/products/primary-antibodies/snap29-antibody-epr91982-ab181151'>ab181151</a>

Lane 2:

Jurkat lysate

Secondary

All lanes:

Peroxidase conjugated Goat anti-Rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 29 kDa

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