Anti-Talin 1 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Talin 1 antibody (AB71333)

IHC image of Talin 1 staining in Human Normal Kidney FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab71333, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Talin 1 antibody (AB71333)

ab71333 staining Talin 1 in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab71333 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Talin 1 antibody (AB71333)

Image : Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

Sample : mouse endoderm cell

Preparation :

Fix in 3% PFA in PBS for 30 min at RT

Primary antibody : Rabbit anti-talin (ab71333), 1 : 100

Secondary antibody : Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Cy3® 488) preadsorbed (ab97075), 1 : 100

Nuclear counterstain : DAPI

Rhodamine-phalloidin, 1 : 100

Nuclei were counterstained with DAPI

Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

• WB

AbReview32678****

Western blot - Anti-Talin 1 antibody (AB71333)

Detection method : Licor System

All lanes:

Western blot - Anti-Talin 1 antibody (ab71333) at 1/1000 dilution

Lane 1:

Human platelets at 20 µg

Lane 2:

Mouse platelets at 20 µg

Secondary

All lanes:

Goat polyclonal to rabbit IgG conjugated to IRDye 800CW (undiluted)

Predicted band size: 269 kDa

false

Exposure time: 1min

This image is courtesy of an anonymous abreview.

• WB

Lab

Western blot - Anti-Talin 1 antibody (AB71333)

Western blot : Anti-TLN1 antibody (ab71333) staining at 1 ug/ml, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab71333 was shown to bind specifically to TLN1. A band was observed at 270 kDa in wild-type PC-3 cell lysates with no signal observed at this size in TLN1 knockout cell line. To generate this image, wild-type and TLN1 knockout PC-3 cell lysates were analysed. First, 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Talin 1 antibody (ab71333) at 1 µg/mL

Lane 1:

Wild-type PC-3 cell lysate at 20 µg

Lane 2:

TLN1 knockout PC-3 cell lysate at 20 µg

Lane 3:

TLN1 knockout PC-3 cell lysate at 10 µg

Lane 4:

HeLa cell lysate at 20 µg

Lane 5:

SK-OV-3 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 270 kDa

false

• WB

Project

Western blot - Anti-Talin 1 antibody (AB71333)

All lanes:

Western blot - Anti-Talin 1 antibody (ab71333) at 1 µg/mL

Lane 1:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 2:

Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 269 kDa

Observed band size: 100 kDa,230 kDa,270 kDa

false

Exposure time: 8min