Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free

• Flow Cyt

Lab

Flow Cytometry - Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free (AB326428)

This data was developed using ab326425, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of bEnd.3 (mouse brain endothelial cell, Right) EL4 (mouse lymphoma T lymphocyte, Left) cells labelling CLEC14A with ab326425 at 1/50 dilution (1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Negative control : EL4

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free (AB326428)

This data was developed using ab326425, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse squamous cell carcinoma tissue labeling CLEC14A with ab326425 at 1/500 (0.994 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse squamous cell carcinoma.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free (AB326428)

This data was developed using ab326425, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) bEnd.3 (mouse brain endothelial cell) cell pellet (B) EL4 (mouse lymphoma T lymphocyte) cell pellet tissue labeling CLEC14A with ab326425 at 1/200 (2.485 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) bEnd.3 cell pellet, no staining on (B) EL4 cell pellet.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free (AB326428)

This data was developed using ab326425, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling CLEC14A with ab326425 at 1/500 (0.994 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse pancreas.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free (AB326428)

This data was developed using ab326425, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling CLEC14A with ab326425 at 1/500 (0.994 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse lung.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free (AB326428)

This data was developed using ab326425, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CLEC14A with ab326425 at 1/500 (0.994 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Weak staining on endothelial cells of mouse cerebrum.

The primary antibody was incubated 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

Lab

Western blot - Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free (AB326428)

This data was developed using ab326425, the same antibody clone in a different buffer formulation.

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

Low expression : liver, brain, skeletal muscle.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CLEC14A antibody [EPR30282-549] (<a href='/ko/products/primary-antibodies/clec14a-antibody-epr30282-549-ab326425'>ab326425</a>) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse skeletal tissue lysate 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: 100 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-CLEC14A antibody [EPR30282-549] - BSA and Azide free (AB326428)

This data was developed using ab326425, the same antibody clone in a different buffer formulation.

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

Negative control : EL4, NIH/3T3.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 21706054).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CLEC14A antibody [EPR30282-549] (<a href='/ko/products/primary-antibodies/clec14a-antibody-epr30282-549-ab326425'>ab326425</a>) at 1/1000 dilution

Lane 1:

bEnd.3 (mouse brain endothelial cell) whole cell lysate at 20 µg

Lane 2:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 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 1/20000 dilution

Observed band size: 100 kDa,36 kDa

false

Exposure time: 59s