Anti-Lamin A + Lamin C antibody [EP4520-16]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EP4520-16] (AB133256)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labelling Lamin A with ab133256 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EP4520-16] (AB133256)

Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labelling Lamin A with ab133256 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Lamin A + Lamin C antibody [EP4520-16] (AB133256)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133256 observed at 70-75 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab133256 was shown to react with LMNA in wild-type HAP1 cells in Western blot. Loss of signal was observed when LMNA knockout sample was used. HAP1 wild-type and LMNA knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween^®) before incubation with ab133256 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Lamin A + Lamin C antibody [EP4520-16] (ab133256) at 1/5000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

LMNA knockout HAP1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 74 kDa

Observed band size: 70-75 kDa

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• WB

Unknown

Western blot - Anti-Lamin A + Lamin C antibody [EP4520-16] (AB133256)

All lanes:

Western blot - Anti-Lamin A + Lamin C antibody [EP4520-16] (ab133256) at 1/10000 dilution

Lane 1:

HeLa treated with staurosporine cell lysate at 10 µg

Lanes 2 and 5:

HeLa cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

HeLa treated with camptothecine cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 74 kDa

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• WB

CiteAb

Western blot - Anti-Lamin A + Lamin C antibody [EP4520-16] (AB133256)

Lamin A + Lamin C western blot using anti-Lamin A + Lamin C antibody [EP4520-16] ab133256. Publication image and figure legend from Zhang, X., Liu, P., et al., 2020, J Cancer, PubMed 31897243.

ab133256 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 ab133256 please see the product overview.

LW6 attenuates the accumulation of cellular YAP1 and its nuclear location. After treating 6606PDA cells (A) or PANC-1 cells (B) with LW6 for 12 hours, LW6 decreased the accumulation of YAP1 when compared to Sham treated cells. In addition, after treating 6606 PDA with LW6 cells for 6 hours, the accumulation of YAP1 in the nucleus was attenuated when compared to Sham-treated cells (C and D). Moreover, after incubation of PANC-1 cells with 80 µM LW6 for 12 hours, LW6 decreased the accumulation of nuclear YAP1 (E). n = 6 per group for A, n =5 per group for B, n = 10 per group for D, n = 2 per group for E. Bar = 5µm. Arrows point at nuclei.

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