Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Immunohistochemical analysis of paraffin-embedded human mammary gland tissue labeling Fibronectin with ab268021 at 1/2000 dilution (0.23 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the stroma of human mammary gland (PMID : 650101). Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Immunohistochemical analysis of paraffin-embedded human esophageal squamous cell carcinoma tissue labeling Fibronectin with ab268021 at 1/2000 dilution (0.23 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the stroma of human esophageal squamous cell carcinoma (PMID : 30314454). Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Immunohistochemical analysis of paraffin-embedded human pancreatic carcinoma labeling Fibronectin with ab268021 at 1/2000 dilution (0.228 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).The section was incubated with ab268021 at 4°C overnight. Positive staining in the human pancreatic carcinoma noted. Counterstained with hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat colon tissue labeling Fibronectin with ab268021 at 1/100 dilution followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Positive staining mainly on smooth muscle of rat colon (PMID : 17881565). The nuclear counterstain is DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

• Flow Cyt

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Flow Cytometry - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Flow cytometric analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling Fibronectin with ab268021 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse colon tissue labeling Fibronectin with ab268021 at 1/100 dilution followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Positive staining mainly on smooth muscle of mouse colon (PMID : 17881565). The nuclear counterstain is DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Fibronectin with ab268021 at 1/2000 dilution (0.23 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the smooth muscle of mouse stomach (PMID : 650101). Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Fibronectin with ab268021 at 1/2000 dilution (0.23 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the perivascular tissue of sinusoids of rat liver (PMID : 650101). Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

• WB

Unknown

Western blot - Anti-Fibronectin antibody [EPR23110-25] - BSA and Azide free (AB268023)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 44 seconds.

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 28367057).

ab268021 was shown to specifically react with Fibronectin in wild-type HAP1 cells as signal was lost in Fibronectin knockout cells. Wild-type and Fibronectin knockout samples were subjected to SDS-PAGE. ab268021 and ab179475 (Rabbit anti-Integrin alpha V loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/5000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument using the ECL technique.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268021).

All lanes:

Western blot - Anti-Fibronectin antibody [EPR23110-25] (<a href='/en-us/products/primary-antibodies/fibronectin-antibody-epr23110-25-ab268021'>ab268021</a>) at 1/1000 dilution

Lane 1:

Human colon lysate at 20 µg

Lane 2:

Human stomach lysate at 20 µg

Lane 3:

Wild-type HAP1 whole cell lysate at 40 µg

Lane 4:

Fibronectin knockout HAP1 whole cell lysate at 40 µg

Secondary

Lanes 1 - 2:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Lanes 3 - 4:

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

Predicted band size: 262 kDa

Observed band size: 285 kDa

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