Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] - BSA and Azide free

• WB

Unknown

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] - BSA and Azide free (AB247992)

This data was developed using ab124764, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] (<a href='/en-us/products/primary-antibodies/interferon-alpha-beta-receptor-1-antibody-epr6244-ab124764'>ab124764</a>) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HeLa cell lysate treated with IFN-alpha at 10 µg

Lane 3:

K562 cell lysate at 10 µg

Lane 4:

U937 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/200 dilution

Predicted band size: 64 kDa

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• WB

Lab

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] - BSA and Azide free (AB247992)

This data was developed using ab124764, the same antibody clone in a different buffer formulation. Western blot : Anti-IFNAR1 antibody [EPR6244] (ab124764) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab124764 was shown to bind specifically to IFNAR1. A band was observed at 116 kDa in wild-type HeLa cell lysates with no signal observed at this size in IFNAR1 knockout cell line. To generate this image, wild-type and IFNAR1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] (<a href='/en-us/products/primary-antibodies/interferon-alpha-beta-receptor-1-antibody-epr6244-ab124764'>ab124764</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human IFNAR1 (Interferon alpha/beta receptor 1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ifnar1-interferon-alpha-beta-receptor-1-knockout-hela-cell-line-ab261782'>ab261782</a>)

Lane 2:

IFNAR1 knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 64 kDa

Observed band size: 116 kDa

false

• WB

Lab

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] - BSA and Azide free (AB247992)

This data was developed using ab124764, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] (<a href='/en-us/products/primary-antibodies/interferon-alpha-beta-receptor-1-antibody-epr6244-ab124764'>ab124764</a>) at 1/1000 dilution

Lane 1:

HEK-293 whole cell lysate at 10 µg

Lane 2:

HeLa whole cell lysate at 10 µg

Lane 3:

LNCaP whole cell lysate at 10 µg

Lane 4:

SH-SY5Y whole cell lysate at 10 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: 64 kDa

Observed band size: 90-130 kDa

false

• WB

Lab

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] - BSA and Azide free (AB247992)

This data was developed using ab124764, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

In mouse and rat tissue lysates this product detects a band in the region of 100 kDa, however we believe this band is non-specific and is not interferon receptor alpha as the immunogen for this antibody shares only 55% homology with the mouse and rat protein. In addition this band also migrates at a lower molecular weight than that detected in human samples.

All lanes:

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] (<a href='/en-us/products/primary-antibodies/interferon-alpha-beta-receptor-1-antibody-epr6244-ab124764'>ab124764</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Rat brain tissue lysate at 10 µg

Predicted band size: 64 kDa

Observed band size: 90 kDa

false

• WB

Lab

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] - BSA and Azide free (AB247992)

This data was developed using ab124764, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

In mouse and rat tissue lysates this product detects a band in the region of 100 kDa, however we believe this band is non-specific and is not interferon receptor alpha as the immunogen for this antibody shares only 55% homology with the mouse and rat protein. In addition this band also migrates at a lower molecular weight than that detected in human samples.

All lanes:

Western blot - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] (<a href='/en-us/products/primary-antibodies/interferon-alpha-beta-receptor-1-antibody-epr6244-ab124764'>ab124764</a>) at 1/1000 dilution

Lane 1:

Mouse brain whole tissue lysate at 10 µg

Lane 2:

Mouse heart whole tissue lysate at 10 µg

Lane 3:

Mouse kidney whole tissue lysate at 10 µg

Lane 4:

Mouse spleen whole tissue lysate at 10 µg

Lane 5:

Rat brain whole tissue lysate at 10 µg

Lane 6:

Rat heart whole cell lysate at 10 µg

Lane 7:

Rat kidney whole cell lysate at 10 µg

Lane 8:

Rat spleen whole tissue lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit IgG (H+L), peroxidase conjugated at 1/2000 dilution

Predicted band size: 64 kDa

false

Exposure time: 3min

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Interferon alpha/beta receptor 1 antibody [EPR6244] - BSA and Azide free (AB247992)

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.