Stressgen, BioMol, Alexis Corporation, Assay Designs, Alexis

antibody

Lamin A/C monoclonal antibody (4C11)

ALX-804-672-C200

200 µg

monoclonal

mouse

nonconjugated

4C11

The same clone is also sold as:

• Abcam: ab238303
• MilliporeSigma: SAB4200236
• Cell Signaling Technology: 4777, 8617

human, mouse, rat

western blot, immunocytochemistry, immunoprecipitation

ALX-804-672-C200

Lamin A/C monoclonal antibody (4C11)

Mouse

Monoclonal

Antibody

200 µg

Figure 2: Immunoprecipitation of lamin A/C using MAb to Lamin A/C (4C11) (Prod. No. ALX-804-672).Method: For each IP 2mg of whole cell lysate from HeLa cells ectopically expressing FLAG-tagged lamin A were used. The lysates were incubated with the indicated volumes of hybridoma supernatant  for 1 hour at 4°C. Then 30µl of Protein A + G sepharose (1:1 suspension) were added and incubated for 1 hour at 4°C. Beads were washed, boiled in 30µl 1x GSD boiling buffer for 5 min, and the proteins were loaded on a 10% SDS-PAG and electrophoretically separated (2% of lysate before and after the IP were also loaded). The proteins were blotted on a nitrocellulose membrane. The membrane was blocked with 3% NFDM-PBS/T for 1 hour and then incubated with MAb to Lamin A/C (4C11) (1:1000 in 0.5% NFDM-PBS/T) over night, 4°C. The membrane was washed 3 x 10 min with PBS/T and then incubated with anti-mouse HRP (1:5000 in 0.5% NFDM-PBS/T) for 1 hour at RT. The membrane was washed 3 x 10 min and then incubated for 1 min with ECL solution. The signal was detected on a HR-HA 30 X-ray film. Exposure time was 5 sec.

Figure 1: Western blot analysis of lamin A/C using MAb to Lamin A/C (4C11) (Prod. No. ALX-804-672). Method: HeLa cells were lysed with 1 x GSD boiling buffer. The cell lysate was electrophoretically separated on a 7.5% SDS-PAG and transferred onto a nitrocellulose membrane. One lane contains approximately 7.5µg of whole cell protein. The nitrocellulose membrane was blocked for 1 h with 3% NFDM/PBS-T and then incubated with MAb to Lamin A/C (4C11) at the indicated dilutions (diluted in 0.5% NFDM/PBS-T) for 2 h at RT. The membrane was washed with PBS-T and then incubated for 1 h with HRP coupled anti-mouse antibody (1:5000). The membrane was washed 3 x 10 min with PBS-T and then incubated for 1 min with ECL solution. The signal was detected on a HR-HA 30 X-ray film. Exposure time was 3 min.

4C11

IgG2a

Human, Mouse, Rat, Monkey, Hamster

Lamin A is a type V intermediate filament protein, involved in maintaining nuclear shape and structure, transcriptional regulation, nuclear pore positioning and function, and heterochromatin organization. Mutations in lamin A are associated with several genetic degenerative disease, including Hutchinson-Gilford progeria syndrome (HGPS), which is caused by Progerin a truncated version of lamin A. A-type lamins have been described prognostic biomarkers for colorectal cancer, and lamin A (but not lamin C) is a potential adult stem cell biomarker in colonic crypts.

Recombinant human lamin A/C (aa 430-545).

Liquid. Hybridoma supernatant containing sodium azide.

ICC, IP, WB

-20°C