Aldolase A Antibody

Western Blot: Aldolase A Antibody [NB600-915]

Western Blot: Aldolase A Antibody [NB600-915] - Analysis using Biotin conjugate of NB600-915. 300 ul aliquots of whole anti-aldolase antiserum used to precipitate varying amounts of purified aldolase & precipitates with controls; compared by SDS-PAGE & Western blot. 1. Purified aldolase 2. 300 ul antiserum with no antigen (negative control) 3. 300 ul antiserum with 100 ul aldolase (2.5 mg/ml) 4. 300 ul antiserum with 200 ul aldolase (2.5 mg/ml) For the precipitation, 300 ul of antiserum & an equal volume of aldolase antigen in PBS incubated 24 hrs at 4C, centrifuged for 6 mins at 13K RPM, washed once with PBS, centrifuged & dissolved in 60 ul 0.1 N NaOH. 90 ul of PBS was added, sample was divided in 2 portions, & an equal volume of reducing (+4% BME) or non-reducing 2X sample buffer added. The reduced samples were boiled for five mins, & all samples were run at 140 V for 45 mins on a 4-20% tris/glycine gradient gel. Gel was stained, destained & imaged.

Western Blot: Aldolase A Antibody [NB600-915]

Western Blot: Aldolase A Antibody [NB600-915] - Figure. Immunoblotting. IgG purified antibody to rabbit muscle aldolase was used at a 1:1000 dilution to detect human aldolase by Western blot. A whole cell lysate prepared from human derived A293 cells was loaded on a 4-12% tris glycine gradient gel for SDS-PAGE. The gel was transferred to nitro-cellulose using standard techniques. Antibody reaction with the membrane occurred overnight at 4 degrees C in TTBS supplemented with 2% non-fat dry milk. Color was allowed to develop using SuperSignal West Pico Chemiluminescent Substrate (PIERCE). Other detection methods will yield similar results. This antibody clearly detects a band at ~41 kDa consistent with human aldolase.

Western Blot: Aldolase A Antibody [NB600-915]

Western Blot: Aldolase A Antibody [NB600-915] - IgG purified antibody to rabbit muscle aldolase was used at a 1:1000 dilution to detect human aldolase by Western blot. A whole cell lysate prepared from human derived A293 cells was loaded on a 4-12% tris glycine gradient gel for SDS-PAGE. The gel was transferred to nitro-cellulose using standard techniques. Antibody reaction with the membrane occurred overnight at 4 C in TTBS supplemented with 2% non-fat dry milk. Color was allowed to develop using SuperSignal West Pico Chemiluminescent Substrate (PIERCE).