Anti-Lamin A抗体

• WB

Lab

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : Lamin A knockout HAP1 cell lysate (20 μg)
Lane 3 : A431 cell lysate (20 μg)
Lane 4 : NIH3T3 cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab26300 observed at 76 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab26300 was shown to recognize Lamin A in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when Lamin A knockout samples were examined. Wild-type and Lamin A knockout samples were subjected to SDS-PAGE. ab26300 1ug/ml and ab8245 (loading control to GAPDH) at a dilution of 1/1000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (ab26300)

Predicted band size: 74 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

ab26300 staining Lamin A in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 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 ab26300 at 5µg/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 magenta). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Project

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

All lanes:

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (ab26300) at 1 µg/mL

All lanes:

Western blot - A-431 whole cell lysate (<a href='/products/cell-lysates/a-431-whole-cell-lysate-ab7909'>ab7909</a>) at 20 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

Predicted band size: 74 kDa

Observed band size: 68 kDa,76 kDa

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• WB

Supplier Data

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

Lysates at 20 µg per lane.

The samples were run on a Bis-Tris gel under reducing conditions.

Western blot : Anti-Lamin A + Lamin C antibody (ab315838) staining at 1/1000 dilution or Anti-Lamin A antibody (ab26300) at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5](ab8245) loading control staining at 1/20000 dilution, shown in red.

In Western blot, ab315838 was shown to bind specifically to Lamin A + Lamin C. Target of interest was observed at 70-75 kDa in wild-type Hela cell lysates (lane 1) with no signal observed at this size in LMNA knockout cell line (lane 2, knockout cell line ab261787/knockout cell lysate ab256979). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

All lanes:

Western blot - Anti-Lamin A + Lamin C antibody [RM1093] (<a href='/products/primary-antibodies/lamin-a-lamin-c-antibody-rm1093-ab315838'>ab315838</a>) at 1/1000 dilution

Lane 1:

Wild-type Hela (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

LMNA knockout Hela whole cell lysate at 20 µg

Lane 3:

HeG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole 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: 70-75 kDa,36 kDa

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• WB

AbReview35406****

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

Blocking : 5% milk for 30 minutes at 22°C

All lanes:

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (ab26300) at 1/1000 dilution

All lanes:

HeLa whole cell extract at 100 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) HRP Conjugate at 1/10000 dilution

Predicted band size: 74 kDa

Observed band size: 76 kDa

true

Exposure time: 15s

This image is courtesy of an anonymous abreview.

• WB

Unknown

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

All lanes:

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (ab26300) at 1 µg/mL

Lane 1:

Western blot - NIH/3T3 whole cell lysate (<a href='/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 10 µg

Lane 2:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 74 kDa

Observed band size: 100 kDa,45 kDa,70 kDa,74 kDa

false

• WB

Unknown

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

All lanes:

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (ab26300) at 1 µg/mL

Lane 1:

Western blot - Recombinant Human Lamin A protein (<a href='/products/proteins-peptides/recombinant-human-lamin-a-protein-ab83472'>ab83472</a>) at 0.1 µg

Lane 2:

Western blot - Recombinant Human Lamin A protein (<a href='/products/proteins-peptides/recombinant-human-lamin-a-protein-ab83472'>ab83472</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 74 kDa

true

Exposure time: 10s

• WB

AbReview38043****

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

All lanes:

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (ab26300) at 1/1000 dilution

Lane 1:

Mouse NIH-3T3 cells - cytosolic fraction at 25 µg

Lane 2:

Mouse NIH-3T3 cells - nuclear fraction at 25 µg

Secondary

All lanes:

HRP conjugated Goat anti-rabbit at 1/5000 dilution

Predicted band size: 74 kDa

Observed band size: 76 kDa

true

Exposure time: 10s

This image is courtesy of an anonymous Abreview

• WB

CiteAb

Western blot - Anti-Lamin A antibody - Nuclear Envelope Marker (AB26300)

Western Blotting using Anti-Lamin A antibody, ab26300. Publication image from Bu, Y. et al., 2016, Nat Commun, 27173017. Legend direct from paper.

PERK-dependent miR-216b induction.(a) PERK+/+ and PERK−/− MEFs were treated with 500 nM TG for indicated times. MiR-216b was assessed by qPCR (left graph); PERK and CHOP were assessed by immunoblot (right). (b) MiR-216b levels were quantified by qPCR following exposure of cells to thapsigargin and a small-molecule PERK inhibitor (left). PERK, eIF2α-p and CHOP induction were assessed by immunoblot (right). (c–e) MEFs of the indicated genotype were treated with TG (500 nM) for indicated intervals. Protein extracts from these cells were immunoblotted for the proteins as indicated (lower panels) and miR-216b levels were quantified by qPCR (upper panels; n=3). (f) CHOP−/− MEFs were transfected with vector or CHOP and 2 days later treated with TG (500 nM) for indicated intervals. Protein extracts from these cells were immunoblotted for CHOP and miR-216b levels quantified by qPCR (n=3). (g) MiR-216b expression and (h) c-Jun mRNA levels were analysed in MMTV-Neu tumours from either PERK+/+ or a PERK−/− background. Data represent mean±s.d. of three independent observations. Statistical significance was analysed analysed by Student's t-test. (*P<0.05, WT versus −/−).

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