Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized bEnd.3 (mouse brain endothelial cell) cells labelling Von Willebrand Factor with ab287962 at 1/100 dilution (5.58 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody used at 1/1000 dilution (2µg/ml) (Green). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5µg/ml)(Red). The Nuclear counterstain was DAPI (Blue). Confocal image showing cytoplasmic staining in bEnd.3 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Colon tissue from wild-type C57BL/6J mice, and (B) Colon tissue from Vwf knockout mice with ab287962 at 1/50 (11.22 μg/ml) dilution. endothelial cells of (A) Colon tissue from wild-type C57BL/6J mice, and no staining on endothelial cells of (B) Colon tissue from Von Willebrand Factor knockout mice. The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Vwf-KO homozygous mice (Strain ID : T028359).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Mouse blood cells cells labelling VonWillebrand Factor with ab287962 at 1/500 dilution (0.1ug)(Right) compared with a Rabbit monoclonal IgG (ab172730) (Left) isotype control. Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab287962. Then stained with anti-CD41 conjugated to APC. Gated on viable cells.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Spleen tissue from wild-type C57BL/6J mice, and (B) Spleen tissue from Vwf knockout mice with ab287962 at 1/50 (11.22 μg/ml) dilution. Positive staining on megakaryocyte of (A) Spleen tissue from wild-type C57BL/6J mice, and no staining on megakaryocyte of (B) Spleen tissue from Von Willebrand Factor knockout mice. The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Vwf-KO homozygous mice (Strain ID : T028359).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Tissue Microarrays stained for Anti-Von Willebrand Factor antibody [EPR25069-131] using ab287962 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab287962 for 30 mins at room temperature followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung (fresh) tissue labeling VonWillebrand Factor with ab287962 at 1/500 (1.11 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on the vascular endothelial cells in rat lung is observed. The nuclear counterstain was DAPI (Blue).

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-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh) tissue labeling VonWillebrand Factor with ab287962 at 1/500 (1.11 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on the endothelial cells in mouse liver is observed. The nuclear counterstain was DAPI (Blue).

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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labelling VonWillebrand Factor with ab287962 at 1/50 (11.1 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection) . Cytoplasmic staining on endothelial cells of rat lung. The section was incubated with ab287962 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh) tissue labeling VonWillebrand Factor with ab287962 at 1/500 (1.11 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on the endothelial cells in rat liver is observed. The nuclear counterstain was DAPI (Blue).

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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling VonWillebrand Factor with ab287962 at 1/50 (11.1 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection) . Cytoplasmic staining on megakaryocyte and platelets of rat spleen. The section was incubated with ab287962 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung (fresh) tissue labeling VonWillebrand Factor with ab287962 at 1/500 (1.11 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on the vascular endothelial cells in mouse lung is observed. The nuclear counterstain was DAPI (Blue).

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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling VonWillebrand Factor with ab287962 at 1/50 (11.1 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Cytoplasmic staining on endothelial cells of mouse colon. The section was incubated with ab287962 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 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-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling VonWillebrand Factor with ab287962 at 1/50 (11.1 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Cytoplasmic staining on megakaryocyte and platelets of mouse spleen. The section was incubated with ab287962 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 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-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tumor tissue labelling VonWillebrand Factor with ab287962 at 1/50 (11.1 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Cytoplasmic staining on endothelial of mouse pancreas tumor. The section was incubated with ab287962 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Lab

Western blot - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

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

In Western blot, Anti-Vinculin antibody [EPR8185] - Loading Control (ab129002) staining at 1/5000 dilution.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Vwf-KO homozygous mice (Strain ID : T028359).

All lanes:

Western blot - Anti-Von Willebrand Factor antibody [EPR25069-131] (<a href='/ko/products/primary-antibodies/von-willebrand-factor-antibody-epr25069-131-ab287962'>ab287962</a>) at 1/1000 dilution

Lane 1:

Wild-type mouse spleen tissue lysate (male) at 20 µg

Lane 2:

VWF knockout mouse spleen tissue lysate (male) at 20 µg

Lane 3:

Wild-type mouse spleen tissue lysate (female) at 20 µg

Lane 4:

VWF knockout mouse spleen tissue lysate (female) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 309 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Von Willebrand Factor antibody [EPR25069-131] - BSA and Azide free (AB287967)

This data was developed using ab287962, the same antibody clone in a different buffer formulation.

Bands below 300kDa were VWF cleavage fragments(PMID : 21909423)

Exposure time : 70 seconds

All lanes:

Western blot - Anti-Von Willebrand Factor antibody [EPR25069-131] (<a href='/ko/products/primary-antibodies/von-willebrand-factor-antibody-epr25069-131-ab287962'>ab287962</a>) at 1/1000 dilution

Lane 1:

Mouse platelet lysate

Lane 2:

Rat platelet lysate

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 309 kDa

false