NBP2-44050-0.5ml

antibody from Novus Biologicals

Targeting: ALDOA

Western blot (Supportive data in Antibodypedia)

ELISA (Data presented on provider website)

Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

NBP2-44050-0.5ml

Provider

Novus Biologicals

Product name

Goat Polyclonal Aldolase A Antibody

Antibody type

Polyclonal

Description

Delipidation and Defibrination.

Reactivity

Human, Rabbit

Host

Goat

Vial size

0.5 ml

Concentration

LYOPH

Storage

Store lyophilized antibody at 4C. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.

Western blot

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: Aldolase A Antibody [NBP2-44050] - Analysis of samples in the image are: 1. Purified aldolase 2. 300 ul antiserum with no antigen (- 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 IP, 300 ul of antiserum and aldolase antigen in PBS was incubated 24 hrs at 4C, centrifuged for 6 minutes at 13K RPM, washed once with PBS, centrifuged and dissolved in 60 ul 0.1 N NaOH. 90 ul of PBS was added, the sample was divided in 2 portions, and an equal volume of reducing or non-reducing 2X sample buffer was added. The reduced samples were boiled for five minutes, and were run at 140 V for 45 minutes on a 4-20% tris/glycine gradient gel. Gel was stained, destained and imaged. Precipitation of aldolase was confirmed by comparison of increase amounts of antigen with control protein by SDS PAGE and observation of a 40-45 kD MW band corresponding to Aldolase. Additional higher and lower molecular weight bands correspond to serum proteins.

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: Aldolase A Antibody [NBP2-44050] - Analysis of human aldolase with IgG purified antibody to rabbit muscle aldolase (NBP2-44050, NB600-915-100ug and NB600-916) at dilution 1:1000. 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 4C in TTBS supplemented with 2% non-fat dry milk. Color was allowed to develop using SuperSignal West Pico Chemiluminescent Substrate. This antibody clearly detects a band at ~41 kDa consistent with human aldolase.

Immunoprecipitation

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Immunoprecipitation: Aldolase A Antibody [NBP2-44050] - Analysis with 300 ul of antiserum and an equal volume of varied amounts of purified aldolase diluted from a stock solution of ~2.5 mg/ml aldolase in PBS. Antibody/Antigen mixture was incubated ~24 hrs at 4C, centrifuged for 6 minutes at 13K RPM, washed once with PBS, centrifuged and dissolved in 60 ul 0.1 N NaOH. 90 ul of PBS was added, the sample was divided in 2 portions, and an equal volume of reducing (+4% BME) or non-reducing 2X sample buffer was added. The reduced samples were boiled for five minutes, and all samples were run at 140 V for ~45 minutes on a 4-20% tris/glycine gradient gel. Gel was stained, destained using standard protocols. Precipitation of aldolase was confirmed by comparison of increasing amounts of antigen with the control protein by SDS PAGE and observation of a 40-45 kD MW band corresponding to Aldolase. Additional higher and lower molecular weight bands correspond to serum proteins.