Anti-Munc18-1 antibody [EPR27956-86]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Munc18-1 with ab315893 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse liver (PMID : 8824310). The section was incubated with ab315893 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh frozen) tissue labeling Munc18-1 with ab315893 at 1/50 (9.8 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal image showing no staining on rat liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab315893 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh frozen) tissue labeling Munc18-1 with ab315893 at 1/50 (9.8 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal image showing no staining on mouse liver (PMID : 8824310). The nuclear counterstain was DAPI (Blue). The section was incubated with ab315893 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Munc18-1 with ab315893 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat kidney. The section was incubated with ab315893 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Munc18-1 with ab315893 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse kidney (PMID : 8824310). The section was incubated with ab315893 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Munc18-1 with ab315893 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat liver. The section was incubated with ab315893 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Munc18-1 with ab315893 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum. The section was incubated with ab315893 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Munc18-1 with ab315893 at 1/100 (4.9 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in rat primary neuron.
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Munc18-1 with ab315893 at 1/100 (0.49 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary neuron.
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocmapus (fresh frozen) tissue labeling Munc18-1 with ab315893 at 1/50 (9.8 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-Munc18-1 (ab315893, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, red) on mouse hippocampus.
Panel B : anti-Munc18-1 stained on mouse hippocampus.
Panel C : anti-NeuN stained in neurons of mouse hippocampus.
Panel D : anti-GFAP stained in astrocytes of mouse hippocampus.
The nuclear counterstain was DAPI (Blue). The section was incubated with ab315893 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocmapus (fresh frozen) tissue labeling Munc18-1 with ab315893 at 1/50 (9.8 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-Munc18-1 (ab315893, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, red) on rat hippocampus.
Panel B : anti-Munc18-1 stained on rat hippocampus.
Panel C : anti-NeuN stained in neurons of rat hippocampus.
Panel D : anti-GFAP stained in astrocytes of rat hippocampus.
The nuclear counterstain was DAPI (Blue). The section was incubated with ab315893 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells labelling Munc18-1 with ab315893 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Munc18-1 with ab315893 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum (PMID : 8824310). The section was incubated with ab315893 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• WB

Supplier Data

Western blot - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

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

Negative control : skeletal muscle (PMID : 7768895).

ab315893 detects both two isoforms of Munc18-1, around 67 and 68 kDa in Western blot (PMID : 20861382).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Munc18-1 antibody [EPR27956-86] (ab315893) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 50 µg

Lane 2:

Human cerebellum tissue lysate at 50 µg

Lane 3:

Human skeletal muscle tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 67 kDa,68 kDa,36 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

ab315893 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 ab315893 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-Munc18-1 antibody [EPR27956-86] (ab315893) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG lysate at 40 µg

Lane 2:

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

false

• WB

Supplier Data

Western blot - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

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

All lanes:

Western blot - Anti-Munc18-1 antibody [EPR27956-86] (ab315893) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 50 µg

Lane 2:

Rat hippocampus tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 67 kDa,68 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

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

Negative control : skeletal muscle (PMID : 7768895).

Low expression : liver (PMID : 7768895).

ab315893 detects both two isoforms of Munc18-1, around 67 and 68 kDa in Western blot (PMID : 20861382).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Munc18-1 antibody [EPR27956-86] (ab315893) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 50 µg

Lane 2:

Mouse cerebellum tissue lysate at 50 µg

Lane 3:

Mouse liver tissue lysate at 50 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 50 µg

Lane 5:

Rat brain tissue lysate at 50 µg

Lane 6:

Rat cerebellum tissue lysate at 50 µg

Lane 7:

Rat liver tissue lysate at 50 µg

Lane 8:

Rat skeletal muscle tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 67 kDa,68 kDa,36 kDa

false

Exposure time: 3s

• Dot

Supplier Data

Dot Blot - Anti-Munc18-1 antibody [EPR27956-86] (AB315893)

Dot blot analysis of Munc18-1 using ab315893 at 1 : 1000 (0.49 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Exposure time : 180 seconds.

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

This antibody does not cross-react with mouse STXBP2.