Anti-CUTC antibody [EPR7929] - BSA and Azide free

• WB

Unknown

Western blot - Anti-CUTC antibody [EPR7929] - BSA and Azide free (AB248644)

This data was developed using ab133762, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CUTC antibody [EPR7929] (<a href='/en-us/products/primary-antibodies/cutc-antibody-epr7929-ab133762'>ab133762</a>) at 1/1000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

Predicted band size: 29 kDa

Observed band size: 29 kDa

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• WB

Lab

Western blot - Anti-CUTC antibody [EPR7929] - BSA and Azide free (AB248644)

This data was developed using ab133762, the same antibody clone in a different buffer formulation.

Lanes 1-5 : Merged signal (red and green). Green - ab133762 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.

ab133762 Anti-CUTC antibody [EPR7929] was shown to specifically react with CUTC in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265860 (knockout cell lysate ab258388) was used. Wild-type and CUTC knockout samples were subjected to SDS-PAGE. ab133762 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-CUTC antibody [EPR7929] (<a href='/en-us/products/primary-antibodies/cutc-antibody-epr7929-ab133762'>ab133762</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CUTC knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CUTC knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cutc-knockout-hela-cell-line-ab265860'>ab265860</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

SH-SY5Y 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

false

• WB

Supplier Data

Western blot - Anti-CUTC antibody [EPR7929] - BSA and Azide free (AB248644)

This data was developed using ab133762, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM /TBST

All lanes:

Western blot - Anti-CUTC antibody [EPR7929] (<a href='/en-us/products/primary-antibodies/cutc-antibody-epr7929-ab133762'>ab133762</a>) at 1/2000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat heart tissue lysate at 20 µg

Lane 7:

Rat kidney tissue lysate at 20 µg

Lane 8:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 29 kDa

Observed band size: 29 kDa

false

Exposure time: 180s

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-CUTC antibody [EPR7929] - BSA and Azide free (AB248644)

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.