Anti-MDH1 antibody [EPR13597(B)] - C-terminal

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (AB180152)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa cells labeling MDH1 with ab180152 at 1/20 dilution followed by Goat anti rabbit IgG (FITC) at 1/75 dilution (red), compared to a Rabbit IgG control (green).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (AB180152)

Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling MDH1 with ab180152 at 1/250 dilution, followed by Goat anti rabbit IgG (Dylight 488) at 1/250 dilution. Counter stained with DAPI.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (AB180152)

ab180152 was shown to react with MDH1 in wild-type HAP1 cells in immunocytochemistry with loss of signal observed in a MDH1 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 ab180152 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.

• IP

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Immunoprecipitation - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (AB180152)

Immunoprecipitation of MDH1 in HAP1 cells. Lysates were prepared and immunoprecipitation was performed using 2 μg of ab180152 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

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:

Immunoprecipitation - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (ab180152) at 2 µg

All lanes:

MDH1 in HAP1 cells at 2 µg

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• IP

Unknown

Immunoprecipitation - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (AB180152)

Western blot analysis of HepG2 cell lysate immunoprecipitated with ab180152 at 1/50 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution

All lanes:

Immunoprecipitation - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (ab180152)

Predicted band size: 36 kDa

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• WB

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Western blot - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (AB180152)

All lanes:

Western blot - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (ab180152) at 1/50000 dilution

Lane 1:

Jurkat cell lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 36 kDa

false

• WB

Supplier Data

Western blot - Anti-MDH1 antibody [EPR13597(B)] - C-terminal (AB180152)

ab180152 was shown to react with MDH1 in wild-type HAP1 cells in Western blot with loss of signal observed in a MDH1 knockout cell line. Wild-type HAP1 and MDH1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab180152 overnight at 4 °C at a 1/10000 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-MDH1 antibody [EPR13597(B)] - C-terminal (ab180152) at 1/10000 dilution

Lane 1:

Wild-type HAP1 lysate at 20 µg

Lane 2:

MDH1 knock-out HAP1 lysate at 20 µg

false