Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] - BSA and Azide free (AB233020)

Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Insulin degrading enzyme / IDE with ab133561 antibody at a dilution of 1/50.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133561).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] - BSA and Azide free (AB233020)

This data was developed using ab133561, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA permeabilized frozen mouse kidney (perfused fixed) tissue labelling Insulin degrading enzyme / IDE with ab133561 at /100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) at 1/1000 dilution.

Confocal image showing positive staining on mouse kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab133561 for 60 mins at room temperature. The section was then mounted using Fluoromount^®. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed ab150081 at 1/1000.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• WB

Supplier Data

Western blot - Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] - BSA and Azide free (AB233020)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : IDE knockout HAP1 cell lysate (20 μg)
Lane 3 : K562 cell lysate (20 μg)
Lane 4 : HepG2 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab133561 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab133561 was shown to specifically react with IDE in wild-type HAP1 cells. No band was observed when IDE knockout samples were examined. Wild-type and IDE knockout samples were subjected to SDS-PAGE. ab133561 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133561).

All lanes:

Western blot - Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] (<a href='/en-us/products/primary-antibodies/insulin-degrading-enzyme-ide-antibody-epr60982-ab133561'>ab133561</a>)

Predicted band size: 118 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] - BSA and Azide free (AB233020)

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.