Anti-IGF1 + IGF2 antibody [EPR5098(2)] - BSA and Azide free

• WB

Unknown

Western blot - Anti-IGF1 + IGF2 antibody [EPR5098(2)] - BSA and Azide free (AB221958)

This data was developed using ab133542, the same antibody clone in a different buffer formulation.

The recombinant IGF-I fragment corresponds to amino acids 49-118 of human IGF-1 with the molecular weight of 10kDa.

All lanes:

Western blot - Anti-IGF1 + IGF2 antibody [EPR5098(2)] (<a href='/en-us/products/primary-antibodies/igf1-igf2-antibody-epr50982-ab133542'>ab133542</a>) at 1/1000 dilution

All lanes:

Recombinant human IGF-I fragment at 0.005 µg

Predicted band size: 22 kDa

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• ELISA

Lab

ELISA - Anti-IGF1 + IGF2 antibody [EPR5098(2)] - BSA and Azide free (AB221958)

This data was developed using ab133542, the same antibody clone in a different buffer formulation.

Primary antibody, ab33542 was used at concentrations between 0.1 - 1000ng/ml. An Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L) was used as the secondary antibody at 1 : 2500 dilution. Antigen concentration was 1000ng/ml.

• WB

Lab

Western blot - Anti-IGF1 + IGF2 antibody [EPR5098(2)] - BSA and Azide free (AB221958)

This data was developed using ab133542, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM /TBST

Predicted band size of IGF1 and IGF2 should be 22kDa, both ab133542 and anti-His tag antibody detected dimer at 44kDa.

ab181602 was used as a GAPDH loading control at 1/1000000 dilution.

All lanes:

Western blot - Anti-IGF1 + IGF2 antibody [EPR5098(2)] (<a href='/en-us/products/primary-antibodies/igf1-igf2-antibody-epr50982-ab133542'>ab133542</a>) at 1/1000 dilution

Lane 1:

HEK-293T transfected with an empty vector whole cell lysate at 20 µg

Lane 2:

HEK-293T transfected with a human IGF1 expression vector containing a His tag whole cell lysate at 20 µg

Lane 3:

HEK-293T transfected with a human IGF2 expression vector containing a His tag whole cell 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

Observed band size: 44 kDa

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Exposure time: 3s

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-IGF1 + IGF2 antibody [EPR5098(2)] - BSA and Azide free (AB221958)

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.