Anti-alpha 1 Antitrypsin antibody [EPR9090]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Antitrypsin antibody [EPR9090] (AB166610)

ab166610 stained HepG2 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab166610 at 1/100 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43μM for 1hour at room temperature.

• WB

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Western blot - Anti-alpha 1 Antitrypsin antibody [EPR9090] (AB166610)

All lanes:

Western blot - Anti-alpha 1 Antitrypsin antibody [EPR9090] (ab166610) at 1/1000 dilution

Lane 1:

Human serum lysate at 10 µg

Lane 2:

Fetal liver lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 46 kDa

Observed band size: 52 kDa

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• WB

Collaborator

Western blot - Anti-alpha 1 Antitrypsin antibody [EPR9090] (AB166610)

ab166610 was shown to react with SERPINA1 in wild-type HepG2 cells in Western blot with loss of signal observed in a SERPINA1 knockout cell line. Wild-type HepG2 and SERPINA1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab166610 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-alpha 1 Antitrypsin antibody [EPR9090] (ab166610) at 1/1000 dilution

Lane 1:

Wild-type HepG2 lysate at 30 µg

Lane 2:

SERPINA1 knock-out HepG2 lysate at 30 µg

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