Anti-Munc18-1 antibody [EPR4850]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Munc18-1 antibody [EPR4850] (AB124920)

ab124920 was shown to react with STXBP1 in wild-type U-87 MG cells in immunocytochemistry with loss of signal observed in STXBP1 knockout cell line ab322392. 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 ab124920 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).

These data were 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Munc18-1 antibody [EPR4850] (AB124920)

ab124920 at 1/250 dilution staining Munc18 in paraffin embedded Human brain tissue by immunohistochemistry

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-Munc18-1 antibody [EPR4850] (AB124920)

Immunoprecipitation of STXBP1 in U-87 MG cells. Lysates were prepared and immunoprecipitation was performed using 2 µg of ab124920 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

These data were 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-Munc18-1 antibody [EPR4850] (ab124920) at 2 µg

All lanes:

U-87 MG cells

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• WB

Unknown

Western blot - Anti-Munc18-1 antibody [EPR4850] (AB124920)

All lanes:

Western blot - Anti-Munc18-1 antibody [EPR4850] (ab124920) at 1/10000 dilution

Lane 1:

SH-SY5Y cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

Y79 cell lysate at 10 µg

Secondary

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HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 68 kDa

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• WB

Lab

Western blot - Anti-Munc18-1 antibody [EPR4850] (AB124920)

ab124920 was shown to react with STXBP1 in wild-type U-87 MG cells in Western blot with loss of signal observed in STXBP1 knockout cell line ab322392. Wild-type U-87 MG and STXBP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab124920 overnight at 4 °C at a 1/10000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were 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-Munc18-1 antibody [EPR4850] (ab124920) at 1/10000 dilution

Lane 1:

Wild-type U-87 MG lysate at 40 µg

Lane 2:

STXBP1 knock-out U-87 MG lysate at 40 µg

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