Goat Anti-Mouse IgG (H+L) Secondary Antibody, Fluoro647 Conjugated

Western blot analysis of KPNB1 using anti-KPNB1 antibody (M01851-2).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human CACO-2 whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human 293T whole cell lysates,
Lane 5: rat NRK whole cell lysates,
Lane 6: mouse NIH/3T3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-KPNB1 antigen affinity purified monoclonal antibody (M01851-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-Fluoro 647 secondary antibody (Catalog # BA1151) at a dilution of 1:2000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for KPNB1 at approximately 97 kDa. The expected band size for KPNB1 is at 97 kDa.

IF analysis of NEFH using anti-NEFH antibody (MA1071).
NEFH was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL mouse anti-NEFH Antibody (MA1071) overnight at 4°C. Fluoro647 Conjugated Goat Anti-Mouse IgG (BA1151) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of NEFH using anti-NEFH antibody (MA1071).
NEFH was detected in a paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL mouse anti-NEFH Antibody (MA1071) overnight at 4°C. Fluoro647 Conjugated Goat Anti-Mouse IgG (BA1151) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.