重组Anti-WSTF抗体[EPR1703]

• WB

Lab

Western blot - Anti-WSTF antibody [EPR1703] (AB109439)

Lanes 1- 2 : Merged signal (red and green). Green - ab109439 observed at 171 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab109439 was shown to react with WSTF in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264907 (knockout cell lysate ab257370) was used. Wild-type HeLa and WSTF knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109439 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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-WSTF antibody [EPR1703] (ab109439) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

WSTF knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BAZ1B (WSTF) knockout HeLa cell line (<a href='/products/cell-lines/human-baz1b-wstf-knockout-hela-cell-line-ab264907'>ab264907</a>)

Predicted band size: 171 kDa

Observed band size: 171 kDa

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• WB

Lab

Western blot - Anti-WSTF antibody [EPR1703] (AB109439)

Lanes 1 - 3 : Merged signal (red and green). Green - ab109439 observed at 171 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab109439 was shown to recognize BAZ1B in wild-type cells along with additional cross-reactive bands as signal was lost in BAZ1B knockout samples. Wild-type and BAZ1B knockout samples were subjected to SDS-PAGE. ab109439 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-WSTF antibody [EPR1703] (ab109439) at 1/1000 dilution

Lane 1:

Wild type HAP1 whole cell lysate at 20 µg

Lane 2:

BAZ1B knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 171 kDa

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• WB

Unknown

Western blot - Anti-WSTF antibody [EPR1703] (AB109439)

All lanes:

Western blot - Anti-WSTF antibody [EPR1703] (ab109439) at 1/1000 dilution

Lane 1:

293T cell lysates at 10 µg

Lane 2:

HeLa cell lysates at 10 µg

Lane 3:

HT-1080 cell lysates at 10 µg

Lane 4:

PC-12 cell lysates at 10 µg

Lane 5:

SH-SY5Y cell lysates at 10 µg

Predicted band size: 171 kDa

Observed band size: 185 kDa

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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-WSTF antibody [EPR1703] (AB109439)

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.