Anti-Kindlin-1 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kindlin-1 antibody (AB68041)

IHC image of Kindlin-1 staining in human liver carcnioma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 ab68041, 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.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Kindlin-1 antibody (AB68041)

ICC/IF image of ab68041 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab68041, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• WB

Project

Western blot - Anti-Kindlin-1 antibody (AB68041)

All lanes:

Western blot - Anti-Kindlin-1 antibody (ab68041) at 1 µg/mL

Lane 1:

Jurkat Whole Cell Lysate - Staurosporine Treated (24hr, 500nM) at 10 µg

Lane 2:

TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate at 10 µg

Lane 3:

Y79 (Human retinoblastoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

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

Lane 5:

Hela Whole Cell Lysate - Staurosporine Treated (24hr, 500nM) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP)

Predicted band size: 77 kDa

Observed band size: 45 kDa,74 kDa,77 kDa

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• WB

CiteAb

Western blot - Anti-Kindlin-1 antibody (AB68041)

Kindlin-1 western blot using anti-Kindlin-1 antibody ab68041. Publication image and figure legend from Patel, H., Stavrou, I., et al., 2016, J Mol Cell Biol, PubMed 26993041.

ab68041 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab68041 please see the product overview.

Kin1 regulates ac-tub levels in an HDAC6-dependent manner. (A) al-tub and ac-tub levels were determined in NT and Kin1 siRNA ± tubacin-treated mitotic cells by quantification of immunofluorescence images. The double thymidine block and release protocol was used to enrich for mitotic cells. (B) ac-tub levels in non-mitotic cells were determined 45 min after disruption of the MT cytoskeleton in MT regrowth assays by quantification of immunofluorescence images in NT or Kin1 siRNA ± tubacin-treated cells. For A and B, >250 cells were measured. (C) ac-tub and al-tub levels determined 0 and 5 h after tubacin washout by quantification of western analysis in cells treated with siRNAs as indicated (top). Representative western blots from which al-tub and ac-tub levels were determined and siRNA depletion confirmed are shown (bottom). In all graphs shown, n ≥ 3, error bars are ±SEM. P-values, t-test, *<0.05, **<0.01, n.s. = not significant.

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