NB600-915-100ug

antibody from Novus Biologicals

Targeting: ALDOA

ELISA

Antibody data

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Validation data

Product number: NB600-915-100ug - Provider product page
Provider: Novus Biologicals
Product name: Goat Polyclonal Aldolase A Antibody
Antibody type: Polyclonal
Description: Ion exchange chromatography. This antibody will detect human Aldolase. Cross reactivity against Aldolase from other tissues and species may also occur. It has been reported that this antibody can detect human Aldolase on immunoblot showing a 41 kDa band in lysates from MCF7, NMB231 and HBL100 cell lines.
Reactivity: Human, Mouse, Rabbit
Host: Goat
Isotype: IgG
Vial size: 100 ug
Concentration: LYOPH
Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

ALDOA protein structure - NB600-915-100ug shown in red.

Supportive validation

Submitted by: Novus Biologicals (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Novus Biologicals (provider)
Main image
Experimental details: Western Blot: Aldolase A Antibody [NB600-915] - 300 ul aliquots of whole anti-aldolase antiserum 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 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 and an equal volume of 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 (+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.

Supportive validation

Submitted by: Novus Biologicals (provider)
Main image
Experimental details: 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).