ECL Substrate Kit (High Sensitivity)

• WB

Unknown

Western blot - ECL Substrate Kit (High Sensitivity) (AB133406)

Western blot comparison 1/25000 anti-Transferrin primary antibody, 1/50000 secondary antibody, 20 second exposure.

Anti-Transferrin antibody (ab1223) at 1/25000 dilution. Lanes 1-14 Transferrin protein (ab91435), Loading dilution (Lanes 1-8) : 12.5, 6.2, 3.1, 1.6, 0.8, 0.3, 0.19μg protein.

Secondary

HRP conjugated polyclonal to Rabbit IgG (ab97080) at 1/50000 developed using Optiblot ECL Detect and Vendor A ECL subtrate.

Exposure time : 20 seconds

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• Dot

Lab

Dot Blot - ECL Substrate Kit (High Sensitivity) (AB133406)

Dot blot of NF-kB p65 peptides, probed with ab19870 at 1 μg ml-1.

Lane 1 : NF-κB p65 (acetyl K310) peptide

Lane 2 : NF-κB p65 (non-acetyl K310) peptide

Lane 3 : NF-κB p65 (acetyl K218) peptide

In dot blot, ab19870 was shown to bind specifically to a NF-κB p65 peptide containing acetyl K310 and not to the non-acetylated version, or to a NF-κB p65 peptide containing acetyl K218.

Nitrocellulose membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibody for 1 h at room temperature. Blots were washed in TBS-T, incubated with secondary antibodies Goat Anti-Rabbit IgG H&L (HRP) at 1/10,000 dilution for 1 h at room temperature, washed again, developed using ECL Substrate Kit ab133406, then imaged.

All lanes:

Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody (<a href='/ko/products/primary-antibodies/nf-kb-p65-acetyl-k310-antibody-ab19870'>ab19870</a>) at 1 µg/mL

Lane 1:

NF-κB p65 (acetyl K310) peptide

Lane 2:

NF-κB p65 (non-acetyl K310) peptide

Lane 3:

NF-κB p65 (acetyl K218) peptide

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (HRP) at 1/10000 dilution

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• Dot

Lab

Dot Blot - ECL Substrate Kit (High Sensitivity) (AB133406)

Dot blot of NF-kB p65 peptides, probed with ab218533 at 1/1000 dilution.

Lane 1 : NF-κB p65 (acetyl K310) peptide

Lane 2 : NF-κB p65 (non-acetyl K310) peptide

Lane 3 : NF-κB p65 (acetyl K218) peptide

In dot blot, ab218533 was shown to bind specifically to a NF-κB p65 peptide containing acetyl K310 and not to the non-acetylated version, or to a NF-κB p65 peptide containing acetyl K218.

Nitrocellulose membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibody for 1 h at room temperature. Blots were washed in TBS-T, incubated with secondary antibodies Goat Anti-Rabbit IgG H&L (HRP) at 1/10,000 dilution for 1 h at room temperature, washed again, developed using ECL Substrate Kit ab133406, then imaged.

All lanes:

Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (<a href='/ko/products/primary-antibodies/nf-kb-p65-acetyl-k310-antibody-epr21781-chip-grade-ab218533'>ab218533</a>) at 1/1000 dilution

Lane 1:

NF-κB p65 (acetyl K310) peptide

Lane 2:

NF-κB p65 (non-acetyl K310) peptide

Lane 3:

NF-κB p65 (acetyl K218) peptide

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (HRP) at 1/10000 dilution

true

• WB

Supplier Data

Western blot - ECL Substrate Kit (High Sensitivity) (AB133406)

Western Blot 1/25000 primary antibody, 1/5000 secondary antibody, 20 second exposure.

Each lane contains the following amount of COX2 recombinant protein (ab58868) : 1) 100ng 2) 50ng 3) 25ng 4) 12.5ng 5) 6.25ng 6) 3.13ng 7) 1.56ng 8) 780pg 9) 390pg 10) 195pg 11) 98pg. The blot was probed with rabbit anti-COX2 antibody (ab15191) at 1/25000 dilution and with a rabbit secondary (ab97080) at 1/5000 dilution. The blot was developed with 20ml of Optiblot ECL Detect.

Exposure time : 20 seconds

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