Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

This data was developed using ab108595 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling Lamin A + Lamin C with ab108595 at a concentration of 0.01µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32 mins. ab108595 anti-Lamin A + Lamin C antibody [EPR4100] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labeling Lamin A + C with ab108595 at 1/250 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Histological analyses of hiPSCs (Human induced pluripotent stem cell) transplanted into the subretinal space of nude rats.

Eye balls were excised from a nude rat 7 weeks after subretinal transplantation with 1×10⁴ hiPSCs. Transplanted tissues were fixed with 4% paraformaldehyde. Paraffin embedded tissue sections were stained with haematoxylin/eosin. Then, the paraffin sections were deparaffinized with xylene and sequential 100%, 95%, 80%, 70% ethanol treatments for 5 min each. The sections were treated with 10 mM citric acid (pH 6) at 95°C for 50 min followed by permeation with 0.4% Triton-X in PBS at room temperature for 30 min.

The deparaffinized sections were stained with ab108595 (Panel M), Hoechst 33258 (Panel N).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

Kanemura et al PLoS One. 2014 Jan 14;9(1):e85336. doi: 10.1371/journal.pone.0085336. eCollection 2014. Fig 6. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunocytochemsitry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Lamin A + C with unpurified ab108595 at 1/250 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labeling Lamin A + C with ab108595 at 1/250 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human urothelial carcinoma tissue labeling Lamin A + C with ab108595.

Green - Lamin A + C.

Red - PI.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling Lamin A + C with purified ab108595 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with hematoxylin.

Negative control using PBS instead of primary antibody (inset).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic carcinoma tissue labeling Lamin A + C with ab108595.

Green - Lamin A + C.

Red - PI.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labeling Lamin A + C with ab108595.

Green - Lamin A + C.

Red - PI.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Lamin A + C with purified ab108595 at 1/110 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody.

Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Lamin A + C (green) with purified ab108595 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control : Primary antibody (1/500) and secondary antibody ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with unpurified ab108595 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab108595, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabeled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labeling Lamin A + C with ab108595.

Green - Lamin A + C.

Red - PI.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

• IP

Lab

Immunoprecipitation - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

ab108595 (purified) at 1/30 immunoprecipitating Lamin A + C in HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

Blocking/Dilution buffer and concentration : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

All lanes:

Immunoprecipitation - Anti-Lamin A + Lamin C antibody [EPR4100] - Nuclear Envelope Marker (<a href='/en-us/products/primary-antibodies/lamin-a-lamin-c-antibody-epr4100-nuclear-envelope-marker-ab108595'>ab108595</a>)

Predicted band size: 74 kDa

Observed band size: 74 kDa

false

• WB

Lab

Western blot - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (AB216074)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108595).

ab108595 was shown to specifically react with Lamin A + C (LMNA) in wild type HAP1 cells. No band was observed when Lamin A + C (LMNA) knockout samples were used. Wild-type and Lamin A + C (LMNA) knockout samples were subjected to SDS-PAGE. The membrane was blocked for an hour using 5% milk before ab108595 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/10000 and 1/20000 dilution respectively. 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Lamin A + Lamin C antibody [EPR4100] - BSA and Azide free (ab216074) at 1/10000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

Lamin A/C knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 74 kDa

Observed band size: 64 kDa,76 kDa

false