Anti-NPY5R antibody [EPR8382] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-NPY5R antibody [EPR8382] - BSA and Azide free (AB248642)

This data was developed using ab133757, the same antibody clone in a different buffer formulation.

Overlay histogram showing SHSY-5Y cells stained with ab133757 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab133757, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 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 >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive result in 4% paraformaldehyde (10 min) fixed SHSY-5Y cells used under the same conditions. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-NPY5R antibody [EPR8382] - BSA and Azide free (AB248642)

This data was developed using ab133757, the same antibody clone in a different buffer formulation.

Immunofluorescent staining of U87 MG cells labelling NPY5R with ab133757 at 1/250.

• WB

Unknown

Western blot - Anti-NPY5R antibody [EPR8382] - BSA and Azide free (AB248642)

This data was developed using ab133757, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-NPY5R antibody [EPR8382] - BSA and Azide free (ab248642) at 1/10000 dilution

Lane 1:

Human hypothalamus tissue lysate at 10 µg

Lane 2:

Human fetal brain tissue lysate at 10 µg

Lane 3:

Ramos cell lysate at 10 µg

Lane 4:

U-87 MG cell lysate at 10 µg

Lane 5:

Caco-2 cell lysate at 10 µg

Lane 6:

BxPC-3 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP-conjugated at 1/2000 dilution

Predicted band size: 51 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-NPY5R antibody [EPR8382] - BSA and Azide free (AB248642)

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.