Anti-Coilin antibody [Pdelta]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Coilin antibody [Pdelta] (AB11822)

ICC/IF image of ab11822 stained MCF-7 cells. The cells were 4% formaldehyde 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 (ab11822, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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

Lab

Western blot - Anti-Coilin antibody [Pdelta] (AB11822)

Western blot : Anti-COIL antibody [Pdelta] (ab11822) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab11822 was shown to bind specifically to COIL. A band was observed at 68 kDa in wild-type HeLa cell lysates with no signal observed at this size in COIL knockout cell line. To generate this image, wild-type and COIL knockout HeLa 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Coilin antibody [Pdelta] (ab11822) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lanes 1 - 4:

Western blot - Human COIL (Coilin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-coil-coilin-knockout-hela-cell-line-ab261757'>ab261757</a>)

Lane 2:

COIL knockout HeLa cell lysate at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 68 kDa

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• WB

CiteAb

Western blot - Anti-Coilin antibody [Pdelta] (AB11822)

Coilin western blot using anti-Coilin antibody [Pdelta] ab11822. Publication image and figure legend from Mahmoudi, S., Henriksson, S., et al., 2010, PLoS Biol, PubMed 21072240.

ab11822 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 ab11822 please see the product overview.

WRAP53 binds coilin and SMN via its WD40 domain and C-terminal region.(A) IP of endogenous WRAP53, coilin, and SMN from U2OS cells followed by immunoblot (IB) with the indicated antibodies. Rabbit and mouse IgG were used as negative controls. (B) Schematic illustration of EGFP-tagged WRAP53 deletion constructs : WRAP53FL (full-length), WRAP53ΔN149+ΔC93 (contains aa 150–455), WRAP53ΔN149 (contains aa 150–584), WRAP53ΔC93 (contains aa 1–455), WRAP53ΔC15 (contains aa 1–533), WRAP53ΔN360 (contains aa 361–548), and WRAP53ΔN449 (contains aa 450–548). The EGFP protein has a molecular weight of approximately 27 kDa. (C) U2OS cells transfected with the indicated WRAP53 constructs for 16 h, followed by IP with GFP antibody. Asterisk indicates the heavy chain. The WRAP53ΔN360 product is 50 kDa in size and is thus covered by the heavy chain.

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