Anti-Von Willebrand Factor antibody [RM1068]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Von Willebrand Factor with ab316979 at 1/2000 (0.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells in human tonsil. The section was incubated with ab316979 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling Von Willebrand Factor with ab316979 at 1/2000 (0.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells in human prostatic hyperplasia. The section was incubated with ab316979 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• Flow Cyt

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Flow Cytometry - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Flow cytometric analysis of Mouse PBMC cells labelling Von Willebrand Factor with ab316979 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 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti-CD41 conjugated to APC.
Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Von Willebrand Factor with ab316979 at 1/2000 (0.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on platelets and megakaryocytes in rat spleen. The section was incubated with ab316979 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling Von Willebrand Factor with ab316979 at 1/2000 (0.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells in rat lung. The section was incubated with ab316979 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Von Willebrand Factor with ab316979 at 1/2000 (0.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on platelets and megakaryocytes in mouse spleen. The section was incubated with ab316979 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung (fresh frozen) tissue labeling Von Willebrand Factor with ab316979 at 1/500 (0.958 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing positive staining in endothelial cells of rat lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab316979 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-Von Willebrand Factor antibody [RM1068] (AB316979)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung (fresh frozen) tissue labeling Von Willebrand Factor with ab316979 at 1/500 (0.958 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing positive staining in endothelial cells of mouse lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab316979 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Von Willebrand Factor with ab316979 at 1/2000 (0.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells in mouse lung. The section was incubated with ab316979 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IP

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Immunoprecipitation - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Von Willebrand Factor was immunoprecipitated from 0.35 mg Rat platelet lysate with ab316979 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316979 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat platelet lysate
Lane 2 : ab316979 IP in Rat platelet lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316979 in rat platelet lysate.

All lanes:

Immunoprecipitation - Anti-Von Willebrand Factor antibody [RM1068] (ab316979) at 1/30 dilution

All lanes:

Rat platelet lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 24s

• IP

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Immunoprecipitation - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Von Willebrand Factor was immunoprecipitated from 0.35 mg Mouse platelet lysate with ab316979 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316979 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse platelet lysate
Lane 2 : ab316979 IP in Mouse platelet lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316979 in mouse platelet lysate.

The bands beneath the target band (300 kDa) are expected to be cleavage fragments (PMID : 21909423).

All lanes:

Immunoprecipitation - Anti-Von Willebrand Factor antibody [RM1068] (ab316979) at 1/30 dilution

All lanes:

Mouse platelet lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 50s

• WB

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Western blot - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

The bands beneath the target band (300 kDa) are expected to be cleavage fragments (PMID : 21909423).

Exposure time : Lanes 1-2 : 37 seconds; Lanes 3-4 : 180 seconds

All lanes:

Western blot - Anti-Von Willebrand Factor antibody [RM1068] (ab316979) at 1/1000 dilution

Lane 1:

Mouse platelet lysate at 2 µg

Lane 2:

Rat platelet lysate at 2 µg

Lane 3:

Human plasma lysate at 40 µg

Lane 4:

Human serum lysate at 40 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 1/100000 dilution

Observed band size: 300 kDa

false

• WB

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Western blot - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

Negative controls : HeLa and HT-29.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Von Willebrand Factor antibody [RM1068] (ab316979) at 1/1000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 40 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 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 1/20000 dilution

Observed band size: 300 kDa,309 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Von Willebrand Factor antibody [RM1068] (AB316979)

The bands beneath the target band (300 kDa) are expected to be cleavage fragments (PMID : 21909423).

Exposure time : Lane 1 : 136 seconds; Lane 2 : 180 seconds

All lanes:

Western blot - Anti-Von Willebrand Factor antibody [RM1068] (ab316979) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

Rat lung tissue lysate at 20 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 1/20000 dilution

Observed band size: 300 kDa

false