Anti-ERGIC3 antibody [EPR8141] - BSA and Azide free

• WB

Unknown

Western blot - Anti-ERGIC3 antibody [EPR8141] - BSA and Azide free (AB248334)

This data was developed using ab129179, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ERGIC3 antibody [EPR8141] (<a href='/en-us/products/primary-antibodies/ergic3-antibody-epr8141-ab129179'>ab129179</a>) at 1/10000 dilution

Lane 1:

HepG2 cell lysate at 10 µg

Lane 2:

MCF7 cell lysate at 10 µg

Lane 3:

293T cell lysate at 10 µg

Lane 4:

K562 cell lysate at 10 µg

Lane 5:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated at 1/2000 dilution

Predicted band size: 43 kDa

Observed band size: 50 kDa

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• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ERGIC3 antibody [EPR8141] - BSA and Azide free (AB248334)

This data was developed using ab129179, the same antibody clone in a different buffer formulation.

ab129179, at 1/250, staining ERGIC3 in HeLa cells by Immunofluorescence.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERGIC3 antibody [EPR8141] - BSA and Azide free (AB248334)

This data was developed using ab129179, the same antibody clone in a different buffer formulation.

ab129179, at 1/100, staining ERGIC3 in paraffin embedded Human brain tissue by Immunohistochemistry.

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

• WB

Lab

Western blot - Anti-ERGIC3 antibody [EPR8141] - BSA and Azide free (AB248334)

This data was developed using ab129179, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab129179 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab129179 Anti-ERGIC3 antibody [EPR8141] was shown to specifically react with ERGIC3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266489 (knockout cell lysate ab257941) was used. Wild-type and ERGIC3 knockout samples were subjected to SDS-PAGE. ab129179 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ERGIC3 antibody [EPR8141] (<a href='/en-us/products/primary-antibodies/ergic3-antibody-epr8141-ab129179'>ab129179</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ERGIC3 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ERGIC3 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ergic3-knockout-hek-293t-cell-line-ab266489'>ab266489</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

MCF-7 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 43 kDa

Observed band size: 50 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-ERGIC3 antibody [EPR8141] - BSA and Azide free (AB248334)

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.