Anti Lamin Immunofluorescence

The pictures were kindly supplied as part of the review submitted by dr josef gotzmann at medical university of vienna austria.

Anti lamin immunofluorescence.

The cells were fixed with 4 paraformaldehyde for 10 minutes permeabilized with 0 1 triton x 100 for 10 minutes and blocked with 1 bsa for 1 hour at room temperature. Immunocytochemistry immunofluorescence anti lamin b1 antibody nuclear envelope marker ab16048 icc if image of ab16048 stained human hela cells. The cleavage of lamins results in nuclear dysregulation and cell death 5 6. Lamin a antibody ma1 06101 in if immunofluorescence analysis of lamin a was performed using 70 confluent log phase hela cells.

For immunofluorescence analysis hela cells were fixed and permeabilized for detection of endogenous lamin b1 using anti lamin b1 recombinant rabbit monoclonal antibody product 702972 1 100 dilution and labeled with goat anti rabbit igg h l superclonal secondary antibody alexa fluor 488 conjugate product a27034 1 2000. This antibody reacts with chicken human mouse rat samples. Anti lamin a c terminal antibody is suitable for use in western blot 0 1 0 2 μg ml using hela nuclear extract. Antibody dilution 1 1 000 2.

During apoptosis lamin a c is specifically cleaved into a large 41 50 kda and a small 28 kda fragment 3 4. 2000 j struct biol 129 313 23. Since both dna blue and lamin a c red are associated with the nuclear compartment this region appears crimson in this image. The cells were methanol fixed 5 min and incubated with the antibody ab16048 1µg ml for 1h at room temperature.

By indirect immunofluorescence the antibody demonstrates specific basement membrane staining of enzymatically unmasked human and animal tissue. Tested in western blot wb immunofluorescence if immunoprecipitation ip and elisa elisa applications. The anti lamin a c antibody reveals strong nuclear lamina staining while anti lamp1 antibody reveals strong cytoplasmic punctate staining of lysosomes and early endosomes. Invitrogen anti lamin b1 monoclonal l 5 catalog 33 2000.

Laminin from engelbreth holm swarm murine sarcoma basement membrane was separated on sds page and probed with rabbit anti laminin ab11575. Antibody dilution 1 2 000 3. Specificity of the anti laminin antibodies is determined by indirect immunofluorescent labeling of formalin fixed paraffin embedded human or animal tissue sections and by dot blot immunoassay. Supplied as 100 µg purified antibody 0 5 mg ml.

Lamin a c is cleaved by caspase 6 and serves as a marker for caspase 6 activation. Immunocytochemistry immunofluorescence anti lamin a antibody 133a2 ab8980 these images show hela cells stained with laminin a antibody ab8980 green and with dapi blue.