Anti-GBP1 antibody [EPR8285] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBP1 antibody [EPR8285] - BSA and Azide free (AB240050)

Immunohistochemical analysis of paraffin embedded Human spleen tissue labelling GBP1 with ab131255 antibody at 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131255).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GBP1 antibody [EPR8285] - BSA and Azide free (AB240050)

Immunofluorescent analysis of IFN-gamma induced HeLa cells labelling GBP1 with ab131255 antibody at 1/100 dilution.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131255).

• WB

Lab

Western blot - Anti-GBP1 antibody [EPR8285] - BSA and Azide free (AB240050)

This data was developed using the same antibody clone in a different buffer formulation (ab131255).

Lanes 1-4 : Merged signal (red and green). Green - ab131255 observed at 68 kDa. Red - loading control ab8245 observed at 36 kDa.

ab131255 Anti-GBP1 antibody [EPR8285] was shown to specifically react with GBP1 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267202 (knockout cell lysate ab257960) was used. Wild-type and GBP1 knockout samples were subjected to SDS-PAGE. ab131255 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-GBP1 antibody [EPR8285] (<a href='/ko/products/primary-antibodies/gbp1-antibody-epr8285-ab131255'>ab131255</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

GBP1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human GBP1 knockout A549 cell line (<a href='/ko/products/cell-lines/human-gbp1-knockout-a549-cell-line-ab267202'>ab267202</a>)

Lane 3:

HeLa treated with 10ng/ml IFN-γ for 24 hours, whole cell lysate at 20 µg

Lane 4:

Untreated HeLa cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 68 kDa

Observed band size: 68 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-GBP1 antibody [EPR8285] - BSA and Azide free (AB240050)

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.