Anti-Aconitase 2 antibody [EPR8283(B)]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aconitase 2 antibody [EPR8283(B)] (AB129105)

ab129105 at 1/100 dilution staining Aconitase 2 in Paraffin-embedded Human kidney tissue by immunohistochemistry.

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

• WB

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Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (AB129105)

All lanes:

Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

K562 cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 85 kDa

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• WB

Lab

Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (AB129105)

Lanes 1 - 2 : Merged signal (red and green). Green - ab129105 observed at 85 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab129105 was shown to specifically react with Aconitase 2 in wild-type HAP1 cells as signal was lost in Hela knockout cells. Wild-type and Hela knockout samples were subjected to SDS-PAGE. ab129105 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105) at 1/1000 dilution

All lanes:

Wild-type HAP1 whole cell lysate at 20 µg

Predicted band size: 37 kDa,85 kDa

Observed band size: 42 kDa,85 kDa

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• WB

AbReview29548****

Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (AB129105)

Blocking : 5% milk for 1 hour at 24°C

All lanes:

Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105) at 1/1000 dilution

Lane 1:

1 month old Mouse Liver Mitochondrial fraction

Lanes 2 - 3:

3 month old Mouse Liver Mitochondrial fraction

Lane 4:

6 month old Mouse Liver Mitochondrial fraction

Secondary

All lanes:

Pig Polyclonal to anti-Rabbit (HRP) at 1/3000 dilution

Predicted band size: 85 kDa

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Exposure time: 1min

This image is courtesy of an abreview submitted by Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom

• OI-RD Scanning

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OI-RD Scanning - Anti-Aconitase 2 antibody [EPR8283(B)] (AB129105)

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.