Anti-SET7 antibody [EPR5574] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-SET7 antibody [EPR5574] - BSA and Azide free (AB239434)

ab124708 (purified) at 1/120 immunoprecipitating SET7 in Jurkat whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) was used for detection at 1/1,500 dilution. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting 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 (ab124708).

All lanes:

Immunoprecipitation - Anti-SET7 antibody [EPR5574] (<a href='/en-us/products/primary-antibodies/set7-antibody-epr5574-ab124708'>ab124708</a>)

Predicted band size: 41 kDa

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• WB

Lab

Western blot - Anti-SET7 antibody [EPR5574] - BSA and Azide free (AB239434)

This data was developed using the same antibody clone in a different buffer formulation (ab124708).

Lanes 1-3 : Merged signal (red and green). Green - ab124708 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab124708 Anti-SET7 antibody [EPR5574] was shown to specifically react with SET7 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265770 (knockout cell lysate ab257668) was used. Wild-type and SET7 knockout samples were subjected to SDS-PAGE. ab124708 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-SET7 antibody [EPR5574] (<a href='/en-us/products/primary-antibodies/set7-antibody-epr5574-ab124708'>ab124708</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SETD7 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SETD7 (SET7) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-setd7-set7-knockout-hela-cell-line-ab265770'>ab265770</a>)

Lane 3:

U-2 OS 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: 41 kDa

Observed band size: 50 kDa

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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-SET7 antibody [EPR5574] - BSA and Azide free (AB239434)

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.