Anti-ATG10 antibody [EPR4804]

• WB

Unknown

Western blot - Anti-ATG10 antibody [EPR4804] (AB124711)

All lanes:

Western blot - Anti-ATG10 antibody [EPR4804] (ab124711) at 1/1000 dilution

Lane 1:

HeLa cell lysates at 10 µg

Lane 2:

K562 cell lysates at 10 µg

Lane 3:

293T cell lysates at 10 µg

Predicted band size: 25 kDa

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• WB

Lab

Western blot - Anti-ATG10 antibody [EPR4804] (AB124711)

Lanes 1-4 : Merged signal (red and green). Green - ab124711 observed at 28 kDa. Red - loading control ab8245 observed at 36 kDa.

ab124711 Anti-ATG10 antibody [EPR4804] was shown to specifically react with ATG10 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265682 (knockout cell lysate ab258786) was used. Wild-type and ATG10 knockout samples were subjected to SDS-PAGE. ab124711 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-ATG10 antibody [EPR4804] (ab124711) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

ATG10 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human ATG10 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-atg10-knockout-hela-cell-line-ab265682'>ab265682</a>)

Lane 3:

K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole 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: 25 kDa

Observed band size: 28 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-ATG10 antibody [EPR4804] (AB124711)

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-ATG10 antibody [EPR4804] (AB124711)

ATG10 western blot using anti-ATG10 antibody [EPR4804] ab124711. Publication image and figure legend from Runwal, G., Stamatakou, E., et al., 2019, Sci Rep, PubMed 31300716.

ab124711 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab124711 please see the product overview.

(a) Representative western blot showing the efficiency of ATG7 and ATG10 knockdown. (b) Representative western blot showing the decrease in LC3-II levels in cells upon ATG7 and ATG10 knockdown. (c) Quantification of the LC3-II levels with respect to levels of GAPDH upon depletion of ATG7 + 10 in HeLa cells. The statistical analysis was performed using one-tailed paired t-test. Error bars represent standard error of the mean (SEM). (d) Representative immunofluorescence images showing the morphology of LC3-positive structures under control and ATG7 + 10 knockdown conditions in HeLa cells. (e,f) Quantification of the LC3-positive structures’ average total area and number under control and ATG7 + 10 knockdown conditions using ImageJ quantification tool (*p < 0.05). Quantification performed from 3 experiments with >20 cells quantified for each condition. Error bars represent standard deviation (SD). Scale bars represent a distance of 10 µm. Full-length blots/gels are presented at the end of the Supplementary File.

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