LS-C744807

antibody from LSBio

Targeting: ALDOA

Western blot (Recommended by provider)

ELISA (Recommended by provider)

Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

LS-C744807

Provider

LSBio

Product name

ALDOA / Aldolase A Antibody (Liquid, Biotin) LS-C744807

Antibody type

Polyclonal

Description

Delipidation, salt fractionation and ion exchange chromatography followed by dialysis.

Reactivity

Human, Rabbit

Host

Goat

Conjugate

Biotin

Isotype

IgG

Storage

Store vial at -20°C or below prior to opening. Dilute 1:10 to minimize loss. Store the vial at -20°C or below after dilution. Avoid freeze-thaw cycles.

Immunoprecipitation

Supportive validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

Anti aldolase antibody– Immunoprecipitation- Immunoprecipitation was performed with 300 ul of anti aldolase antiserum and an equal volume of varied amounts (diluted from a stock solution of ~2.5 mg/ml) of purified aldolase in PBS.  Antibody/Antigen mixture was incubated ~24 hrs at 4°C, 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 and imaged(see attached) 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.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

Anti aldolase antibody – Immunoprecipitation and Western Blot. 300 µl aliquots of whole anti-aldolase antiserum (100-1141) were used to precipitate varying amounts of purified aldolase and precipitates with controls were compared by SDS-PAGE and Western blot. Samples shown in the image are: 1. Purified aldolase 2. 300 µl antiserum with no antigen (negative control) 3. 300 µl antiserum with ~100 µl aldolase (2.5 mg/ml) 4. 300 µl antiserum with ~200 µl aldolase (2.5 mg/ml) For the precipitation, 300 ul of antiserum and an equal volume of aldolase antigen in PBS was incubated ~24 hrs at 4°C, 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 and imaged(see attached) 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.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

Anti aldolase antibody– Immunoprecipitation- Immunoprecipitation was performed with 300 ul of anti aldolase antiserum and an equal volume of varied amounts (diluted from a stock solution of ~2.5 mg/ml) of purified aldolase in PBS.  Antibody/Antigen mixture was incubated ~24 hrs at 4°C, 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 and imaged(see attached) 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.