重组Anti-Tenascin C抗体[EPR4219] - BSA and Azide free (ab271877)

All lanes : Anti-Tenascin C antibody [EPR4219] (ab108930) at 1/500 dilution

Lane 1 : Wild-type U-2 OS BFA (5 ug/mL, 6 h) cell lysate at 40 µg
Lane 2 : Wild-type U-2 OS BFA (0 ug/mL, 6 h) cell lysate at 40 µg
Lane 3 : TNC knockout U-2 OS BFA (5 ug/mL, 6 h) cell lysate at 40 µg
Lane 4 : TNC knockout U-2 OS BFA (0 ug/mL, 6 h) cell lysate at 40 µg
Lane 5 : U-87 MG Treated BFA (5 ug/mL, 6 h) cell lysate at 10 µg
Lane 6 : U-87 MG Control BFA (0 ug/mL, 6 h) cell lysate at 10 µg

Performed under reducing conditions.

Predicted band size: 241 kDa
Observed band size: 122 kDa why is the actual band size different from the predicted?

Western blot: Anti-TNC antibody [EPR4219] (ab108930) staining at 1/500 dilution, shown in black; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab108930 was shown to bind specifically to TNC. A band was observed at 122 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in TNC knockout cell line. To generate this image, wild-type and TNC knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 2 % BSA 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 before development with a high-sensitivity ECL substrate kit and imaged with 5 seconds exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebellar cortex labeling Tenascin C with ab108930 at 1/500 dilution (0.854 μg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA Buffer, pH 9 (ab93684). A ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used. Hematoxylin counterstain. Staining on the molecular layer of rat cerebellar cortex (PMID: 1372043) is observed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108930).

Immunohistochemistry (Frozen sections) analysis of mouse E14 spinal cord labeling Tenascin C with ab108930 at 1/100 dilution (4.27μg/ml). Tissue was fixed with 4% PFA and permeabilized with 0.2% TritonX-100. Antigen retrieval was performed using a heated citrate solution (10mM citrate PH 6.0 + 0.05% Tween-20). ab150077, an AlexaFluor^® 488 Goat anti-Rabbit secondary antibody was used at 1/1000 (2 μg/ml). DAPI nuclear counterstain.

Positive staining on mesenchymal condensations during chondrogenesis of mouse E14 embryo (PMID: 9822997; PMID: 19586317; PMID: 24778247) is observed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108930).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse E14 spinal cord tissue sections labeling Tenascin C with ab108930 at 1/500 dilution (0.854 μg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9 (ab93684). A ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used. Hematoxylin counterstain.

Positive staining on mesenchymal condensations during chondrogenesis of mouse E14 embryo (PMID: 9822997; PMID: 19586317; PMID: 24778247) is observed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108930).

Immunohistochemistry (Frozen sections) analysis of rat cerebellar cortex labeling Tenascin C with ab108930 at 1/100 dilution (4.27μg/ml). Tissue was fixed with 4% PFA and permeabilized with 0.2% TritonX-100. Antigen retrieval was performed using a heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). ab150077, an AlexaFluor^® 488 Goat anti-Rabbit secondary antibody was used at 1/1000 (2 μg/ml). DAPI nuclear counterstain.

Positive staining on the molecular layer of rat cerebellar cortex (PMID: 1372043) is observed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108930).

Immunohistochemistry (Frozen sections) analysis of mouse E14 cerebellar cortex labeling Tenascin C with ab108930 at 1/100 dilution (4.27μg/ml). Tissue was fixed with 4% PFA and permeabilized with 0.2% TritonX-100. Antigen retrieval was performed using a heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). ab150077, an AlexaFluor^® 488 Goat anti-Rabbit secondary antibody was used at 1/1000 (2 μg/ml). DAPI nuclear counterstain.

Positive staining on the molecular layer of mouse E14 cerebellar cortex (PMID: 1372043) is observed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108930).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebellar cortex labeling Tenascin C with ab108930 at 1/500 dilution (0.854 μg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9 (ab93684). Hematoxylin was used to counterstain. A ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used. Staining on the molecular layer of mouse cerebellar cortex (PMID: 1372043) is observed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108930).