Anti-PGD antibody [EPR6565]

• WB

Lab

Western blot - Anti-PGD antibody [EPR6565] (AB129199)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PGD antibody [EPR6565] (ab129199) at 1/5000 dilution

Lane 1:

Mouse brain tissue lysates at 20 µg

Lane 2:

Mouse heart tissue lysates at 20 µg

Lane 3:

Mouse kidney tissue lysates at 20 µg

Lane 4:

Mouse spleen tissue lysates at 20 µg

Lane 5:

Rat brain tissue lysates at 20 µg

Lane 6:

Rat heart tissue lysates at 20 µg

Lane 7:

Rat kidney tissue lysates at 20 µg

Lane 8:

Rat spleen tissue lysates 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

Predicted band size: 53 kDa

Observed band size: 45 kDa

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Exposure time: 3min

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PGD antibody [EPR6565] (AB129199)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PGD with ab129199 at 1/500. Cells were fixed with 100% methanol. ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Control : PBS only.
Nuclear counter stain : DAPI.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PGD antibody [EPR6565] (AB129199)

Overlay histogram showing HEK293 cells stained with ab129199 (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 (ab129199 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 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 >5000 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-PGD antibody [EPR6565] (AB129199)

All lanes:

Western blot - Anti-PGD antibody [EPR6565] (ab129199) at 1/1000 dilution

Lane 1:

Raji cell lysate at 10 µg

Lane 2:

Molt-4 cell lysate at 10 µg

Lane 3:

293T (Human embryonic kidney epithelial cell) cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 53 kDa

Observed band size: 45 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-PGD antibody [EPR6565] (AB129199)

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.