Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free

11 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/2000 dilution (0.29 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the smooth muscle of rat 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 (Anti-Fibronectin antibody [EPR23110-46] ab268020).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/2000 dilution (0.29 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 (Anti-Fibronectin antibody [EPR23110-46] ab268020).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunohistochemical analysis of paraffin-embedded human esophageal squamous cell carcinoma tissue labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/2000 dilution (0.29 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 (Anti-Fibronectin antibody [EPR23110-46] ab268020).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunohistochemical analysis of paraffin-embedded human mammary gland tissue labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/2000 dilution (0.29 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 (Anti-Fibronectin antibody [EPR23110-46] ab268020).

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  Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cells is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Fibronectin antibody [EPR23110-46] ab268020).

• 

  Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat colon tissue labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Positive staining mainly on smooth muscle of rat colon (PMID: 17881565) is observed. The nuclear counterstain is DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary 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 (Anti-Fibronectin antibody [EPR23110-46] ab268020).

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  Western blot - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Fibronectin antibody [EPR23110-46] ab268020).
  

  Anti-Fibronectin antibody [EPR23110-46] ab268020 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. Anti-Fibronectin antibody [EPR23110-46] ab268020 and Anti-Vinculin antibody [EPR8185] ab129002 (Rabbit anti-Vinculin 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 (Goat Anti-Rabbit IgG H&L (HRP) 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.

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

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

  All lanes: Western blot - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022) at 0.585 µg/mL

  Lane 1: Human stomach tissue lysate at 10 µg

  Lane 2: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

  Lane 3: Fibronectin-1 knockout HAP1 whole cell lysate at 40 µg

  Lane 4: Wild-type HAP1 whole cell lysate at 40 µg

  Developed using the ECL technique.

  Predicted band size: 262 kDa

  Observed band size: 285 kDa

• 

  Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse colon tissue labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Positive staining mainly on smooth muscle of mouse colon (PMID: 17881565) is observed. The nuclear counterstain is DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary 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 (Anti-Fibronectin antibody [EPR23110-46] ab268020).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cells is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Fibronectin antibody [EPR23110-46] ab268020).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  Immunohistochemical analysis of paraffin-embedded Human pancreatic carcinoma tissue labeling Fibronectin with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 1/4000 dilution (0.146 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). The section was incubated with Anti-Fibronectin antibody [EPR23110-46] ab268020 at 4°C overnight. Positive staining in the human pancreatic carcinoma counterstained with hematoxylin.
  
  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP)(Goat Anti-Rabbit IgG H&L (HRP polymer) ab214880)
  
  Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

• 

  Western blot - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Fibronectin antibody [EPR23110-46] ab268020).

  Western blot: Rabbit Monoclonal[EPR23110-46] to Fibronectin Anti-Fibronectin antibody [EPR23110-46] ab268020 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 238-268 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in FN1 knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

  All lanes: Western blot - Anti-Fibronectin antibody [EPR23110-46] (Anti-Fibronectin antibody [EPR23110-46] ab268020) at 1/1000 dilution

  Lane 1: Wild-type MCF7 at 20 µg

  Lane 2: Western blot - Human FN1 knockout MCF7 cell line (Human FN1 knockout MCF7 cell line ab286324) at 20 µg

  Lane 3: HepG2 at 20 µg

  Lane 4: PC-3 at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 272 kDa

  Observed band size: 238 kDa, 268 kDa