Anti-ECM Isoform 1 antibody [EPR25810-46] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ECM Isoform 1 antibody [EPR25810-46] - BSA and Azide free (AB314894)

This data was developed using ab314893, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labeling ECM Isoform 1 with ab314893 at 1/5000 (0.104 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human cardiac muscle. The section was incubated with ab314893 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-ECM Isoform 1 antibody [EPR25810-46] - BSA and Azide free (AB314894)

This data was developed using ab314893, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling ECM Isoform 1 with ab314893 at 1/5000 (0.104 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on endothelial cells and platelets in human liver. The section was incubated with ab314893 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-ECM Isoform 1 antibody [EPR25810-46] - BSA and Azide free (AB314894)

This data was developed using ab314893, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling ECM Isoform 1 with ab314893 at 1/5000 (0.104 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on platelets in human spleen. The section was incubated with ab314893 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

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ECM Isoform 1 antibody [EPR25810-46] - BSA and Azide free (AB314894)

This data was developed using ab314893, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling ECM Isoform 1 with ab314893 at 1/100 (1.044 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 HUVEC cell line. Negative control : A549. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ 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.

• WB

Supplier Data

Western blot - Anti-ECM Isoform 1 antibody [EPR25810-46] - BSA and Azide free (AB314894)

This data was developed using ab314893, the same antibody clone in a different buffer formulation.

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

Low expression : plasma

The molecular weight observed is consistent with what has been described in the literature (PMID : 33179420, 19132231).

All lanes:

Western blot - Anti-ECM Isoform 1 antibody [EPR25810-46] (<a href='/en-us/products/primary-antibodies/ecm-isoform-1-antibody-epr25810-46-ab314893'>ab314893</a>) at 1/1000 dilution

Lane 1:

Human serum at 20 µg

Lane 2:

Human plasma at 20 µg

Secondary

All lanes:

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

Observed band size: 186 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-ECM Isoform 1 antibody [EPR25810-46] - BSA and Azide free (AB314894)

This data was developed using ab314893, the same antibody clone in a different buffer formulation.

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

Negative control : A549

The molecular weight observed is consistent with what has been described in the literature (PMID : 33179420, 19132231).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-ECM Isoform 1 antibody [EPR25810-46] (<a href='/en-us/products/primary-antibodies/ecm-isoform-1-antibody-epr25810-46-ab314893'>ab314893</a>) at 1/1000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma epithelial cell) whole cell 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 1/100000 dilution

Observed band size: 186 kDa,36 kDa

false

Exposure time: 180s