Anti-PARK7/DJ1 antibody [EP2816Y]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PARK7/DJ1 antibody [EP2816Y] (AB76241)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling PARK7/DJ1 with ab76241 at 1/20 dilution (7.5 ug/mL) (Red) compared with a isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-PARK7/DJ1 antibody [EP2816Y] (AB76241)

ab76241 was shown to react with PARK7 in wild-type HEK-293T cells in immunocytochemistry with loss of signal observed in PARK7 knockout cell line ab266338. 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 ab76241 at 1/100 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 µg/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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARK7/DJ1 antibody [EP2816Y] (AB76241)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling PARK7/DJ1 with ab76241 at 1/500 (0.494 ug/ml) dilution.

Positive staining on human cerebrum tissue. The section was incubated with ab76241 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARK7/DJ1 antibody [EP2816Y] (AB76241)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling PARK7/DJ1 with ab76241 at 1/500 (0.494 ug/ml) dilution.

Positive staining on mouse cerebrum tissue. The section was incubated with ab76241 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARK7/DJ1 antibody [EP2816Y] (AB76241)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling PARK7/DJ1 with ab76241 at 1/500 (0.494 ug/ml) dilution.

Positive staining on rat cerebrum tissue. The section was incubated with ab76241 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Collaborator

Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (AB76241)

ab76241 was shown to react with PARK7 in wild-type HEK-293T cells in Western blot with loss of signal observed in PARK7 knockout cell line ab266338. Wild-type HEK-293T and PARK7 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab76241 overnight at 4 °C at a 1/2000 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-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/2000 dilution

Lane 1:

Wild-type HEK-293T lysate at 30 µg

Lane 2:

PARK7 knock-out HEK-293T lysate at 30 µg

Lane 2:

Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 30 µg

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• WB

Lab

Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (AB76241)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time :
Lane 1 : 10 seconds;
Lane 2 : 5 seconds;
Lane 3 : 3 seconds.

All lanes:

Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/2000 dilution

Lane 1:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 15 µg

Lane 2:

Mouse brain lysate at 15 µg

Lane 3:

Rat brain lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/200000 dilution

Predicted band size: 20 kDa

Observed band size: 24 kDa

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