Anti-SNAP29 antibody [EPR9199] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-SNAP29 antibody [EPR9199] - BSA and Azide free (AB240110)

ab138500 (purified) at 1/20 immunoprecipitating SNAP29 in 10 μg HeLa cell lysate (Lanes 1 and 2, observed at 29 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution. Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab138500).

All lanes:

Immunoprecipitation - Anti-SNAP29 antibody [EPR9199] (<a href='/en-us/products/primary-antibodies/snap29-antibody-epr9199-ab138500'>ab138500</a>)

Predicted band size: 21 kDa,29 kDa

Observed band size: 21 kDa,22 kDa

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• WB

Lab

Western blot - Anti-SNAP29 antibody [EPR9199] - BSA and Azide free (AB240110)

This data was developed using the same antibody clone in a different buffer formulation (ab138500).

Lanes 1-3 : Merged signal (red and green). Green - ab138500 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.

ab138500 Anti-SNAP29 antibody [EPR9199] 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. ab138500 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 [EPR9199] (<a href='/en-us/products/primary-antibodies/snap29-antibody-epr9199-ab138500'>ab138500</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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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-SNAP29 antibody [EPR9199] - BSA and Azide free (AB240110)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.