Anti-GBA antibody [EPR26755-29] - BSA and Azide free (Capture)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GBA antibody [EPR26755-29] - BSA and Azide free (Capture) (AB309228)

ab309228 was shown to react with GBA1 in wild-type RPE-1 cells in immunocytochemistry with loss of signal observed in a GBA1 knockout cell line. Wild-type and knockout cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab309228 at 1/500 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 ug/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

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.

• sELISA

Supplier Data

Sandwich ELISA - Anti-GBA antibody [EPR26755-29] - BSA and Azide free (Capture) (AB309228)

Sandwich ELISA of ab309266 with the capture antibody dilution at 2 µg/mL and detector antibody dilution at 0.5 µg/mL.

• WB

Supplier Data

Western blot - Anti-GBA antibody [EPR26755-29] - BSA and Azide free (Capture) (AB309228)

ab309228 was shown to react with GBA1 in wild-type HAP1 cells in Western blot with loss of signal observed in a GBA1 knockout cell line. Wild-type HAP1 and GBA1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab309228 overnight at 4 °C at a 1/200 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-GBA antibody [EPR26755-29] - BSA and Azide free (Capture) (ab309228) at 1/200 dilution

Lane 1:

Wild-type HAP1 lysate at 30 µg

Lane 2:

GBA1 knock-out HAP1 lysate at 30 µg

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