Anti-GABARAPL2/GATE-16 antibody [EP4808]

• WB

Lab

Western blot - Anti-GABARAPL2/GATE-16 antibody [EP4808] (AB126607)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : GABARAPL2 / GATE-16 knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : Human tonsil lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab126607 observed at 15 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab126607 was shown to specifically react with GABARAPL2 / GATE-16 when GABARAPL2 / GATE-16 knockout samples were used. Wild-type and GABARAPL2 / GATE-16 knockout samples were subjected to SDS-PAGE. ab126607 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-GABARAPL2/GATE-16 antibody [EP4808] (ab126607)

Predicted band size: 14 kDa

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• WB

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Western blot - Anti-GABARAPL2/GATE-16 antibody [EP4808] (AB126607)

All lanes:

Western blot - Anti-GABARAPL2/GATE-16 antibody [EP4808] (ab126607) at 1/1000 dilution

Lane 1:

Human fetal brain tissue lysate at 10 µg

Lane 2:

Human heart tissue lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Lane 4:

Human spleen tissue lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-GABARAPL2/GATE-16 antibody [EP4808] (AB126607)

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.

• WB

CiteAb

Western blot - Anti-GABARAPL2/GATE-16 antibody [EP4808] (AB126607)

Western Blotting using Anti-GABARAPL2/GATE-16 antibody [EP4808], ab126607. Publication image from Holdgaard, S. G. et al., 2019, Nat Commun, 31519908. Legend direct from paper.

PCM1 accumulation and mitotic abnormalities show GABARAP selectivity. a Immunoblot of U2OS cell extracts following depletion of LC3B, GABARAP and GABARAPL2 individually or in combination as indicated showing levels of PCM1. GAPDH is used as loading control. b Densitometric quantification of PCM1 levels relative to GAPDH, represented in (a). Columns represent the mean ± SD, n = 3, ns > 0.05, **P ≤ 0.01, ***P ≤ 0.001. Unpaired Student’s t-test, two-tailed. c Representative images of PCM1-stained CS in U2OS cells depleted of the denoted ATG8 proteins. d Representative images of mitotic abnormalities in U2OS cells stained for γ-tubulin, β-tubulin and Hoechst33342 following depletion of the denoted ATG8 proteins. e Quantification of phenotype distribution in (d). Columns represent the mean ± SD, n = 3 of ≥20 cells, ns P > 0.05, **P ≤ 0.01. Unpaired Student’s t-test, two-tailed. Abnormal refers to cells exhibiting multipolar, diffuse bipolar or monopolar mitoses. Scale bars, 10 µm. Source data are provided as a Source Data file

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