Anti-AS160 (phospho T642) antibody [EPR2733(2)] - BSA and Azide free

• WB

Lab

Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] - BSA and Azide free (AB218844)

This data was developed using ab131214, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (<a href='/en-us/products/primary-antibodies/as160-phospho-t642-antibody-epr27332-ab131214'>ab131214</a>) at 1.12 µg/mL

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) grown in serum free media overnight whole cell lysate at 10 µg

Lane 2:

HEK-293 grown in serum free media overnight, then treated with 100nM Calyculin A (<a href='/en-us/products/biochemicals/calyculin-a-protein-phosphatase-inhibitor-ab141784'>ab141784</a>) for 50min and then 100ng/ml Insulin was added for the last 20min, whole cell lysate at 10 µg

Lane 3:

HEK-293 grown in serum free media overnight, then treated with 100nM Calyculin A (<a href='/en-us/products/biochemicals/calyculin-a-protein-phosphatase-inhibitor-ab141784'>ab141784</a>) for 50min and then 100ng/ml Insulin was added for the last 20min, whole cell lysate. Then the memb at 10 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: 146 kDa

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• WB

Unknown

Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] - BSA and Azide free (AB218844)

This data was developed using ab131214, the same antibody clone in a different buffer formulation.

Bottom pannel shows detection of total AS160 with a general anti AS160 antibody.

All lanes:

Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (<a href='/en-us/products/primary-antibodies/as160-phospho-t642-antibody-epr27332-ab131214'>ab131214</a>) at 1/1000 dilution

Lane 1:

293T cell lysate - untreated

Lane 2:

293T cell lysate - insulin-treated

Predicted band size: 146 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-AS160 (phospho T642) antibody [EPR2733(2)] - BSA and Azide free (AB218844)

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.

• Dot

Lab

Dot Blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] - BSA and Azide free (AB218844)

This data was developed using ab131214, the same antibody clone in a different buffer formulation.

Dot blot analysis of AS160 (phospho T642) phospho peptide (Lane 1) and AS160 non-phospho peptide (Lane 2) labelling AS160 (phospho T642) phospho peptide with ab131214 at a dilution of 1 : 1000 dilution (1.12μg/ml). A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1 : 20,000 dilution.

Blocking and dilution buffer : 5% NFDM/TBST.