Anti-Talin 1 antibody [97H6] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Talin 1 antibody [97H6] - BSA and Azide free (AB256125)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Talin 1 with ab108480 at 1/5000 dilution (0.241μg/ml) followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). Cytoplasmic staining on mouse spleen. The section was incubated with ab108480 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 Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108480).

• IP

Unknown

Immunoprecipitation - Anti-Talin 1 antibody [97H6] - BSA and Azide free (AB256125)

Talin 1 was immunoprecipitated from NIH/3T3 (mouse embryonic fibroblast), whole cell lysate with ab108480 at 1/30 dilution (2μg in 0.35mg lysates) (16.033μg/ml). Western blot was performed on the immunoprecipitate using ab108480 at 1/1000 dilution (0.481μg/ml). Mouse IgG for IP (HRP) (ab131368) was used at 1/1000 dilution.

Lane 1 : NIH/3T3 whole cell lysate 10μg.

Lane 2 : ab108480 IP in NIH/3T3 whole cell lysate.

Lane 3 : Mouse monoclonal IgG1 (ab18443) instead of ab108480 in NIH/3T3 whole cell lysate.

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

Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108480).

All lanes:

Immunoprecipitation - Anti-Talin 1 antibody [97H6] (<a href='/en-us/products/primary-antibodies/talin-1-antibody-97h6-ab108480'>ab108480</a>)

Predicted band size: 269 kDa

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• WB

Lab

Western blot - Anti-Talin 1 antibody [97H6] - BSA and Azide free (AB256125)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108480).

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 22306379 and 25751747).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Talin 1 antibody [97H6] (<a href='/en-us/products/primary-antibodies/talin-1-antibody-97h6-ab108480'>ab108480</a>) at 0.481 µg/mL

Lane 1:

HUVEC (human umbilical vein endothelial cell), whole cell lysate at 10 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 4:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Lane 5:

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

Lane 6:

MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Predicted band size: 269 kDa

false

Exposure time: 3s