Anti-Exportin-5 antibody [EPR8452] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Exportin-5 antibody [EPR8452] - BSA and Azide free (AB248256)

This data was developed using ab129006, the same antibody clone in a different buffer formulation.

ab129006, at 1/250 dilution, staining Exportin-5 in paraffin-embedded Human colon tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Exportin-5 antibody [EPR8452] - BSA and Azide free (AB248256)

This data was developed using ab129006, the same antibody clone in a different buffer formulation.

Overlay histogram showing HEK293 cells stained with ab129006 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab129006, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• WB

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Western blot - Anti-Exportin-5 antibody [EPR8452] - BSA and Azide free (AB248256)

This data was developed using ab129006, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Exportin-5 antibody [EPR8452] (<a href='/en-us/products/primary-antibodies/exportin-5-antibody-epr8452-ab129006'>ab129006</a>) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

K562 cell lysate at 10 µg

Lane 3:

Human fetal heart tissue lysate at 10 µg

Lane 4:

Human fetal brain tissue lysate at 10 µg

Lane 5:

C6 cell lysate at 10 µg

Lane 6:

RAW 264.7 cell lysate at 10 µg

Lane 7:

PC-12 cell lysate at 10 µg

Lane 8:

NIH/3T3 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated at 1/2000 dilution

Predicted band size: 136 kDa

Observed band size: 136 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-Exportin-5 antibody [EPR8452] - BSA and Azide free (AB248256)

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.