Aldolase A Antibody [Biotin]

Western Blot: Aldolase A Antibody [Biotin] [NB100-2003] - 300 ul aliquots of whole anti-aldolase antiserum were used to precipitate purified aldolase and precipitates with controls were compared by SDS-PAGE and Western blot. Samples shown are: 1. Purified aldolase 2. 300 ul antiserum, no antigen (neg 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 precipitation, 300 ul of antiserum and an equal volume of aldolase antigen in PBS was incubated 24 hrs at 4C, centrifuged 6 minutes at 13K RPM, washed once with PBS, centrifuged and dissolved in 60 ul 0.1 N NaOH. 90 ul of PBS 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, de-stained and imaged(see attached) using standard protocols.

Western Blot: Aldolase A Antibody [Biotin] [NB100-2003] - 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 ug/ml) of purified aldolase in PBS. 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.

Product Discontinued

Aldolase A Antibody [Biotin] Summary

• Immunogen: Aldolase A Antibody [Rabbit Muscle] (Uniprot: P00883)
• Localization: Cytoplasmic
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: ALDOA
• Purity: Whole Antiserum

Applications/Dilutions

• Dilutions:
  • ELISA 1:5000-1:20000
  • Immunoprecipitation 1:100
  • Western Blot 1:500-1:5000
• Application Notes: This antibody has been assayed against 1.0 ug of Aldolase A in a standard capture ELISA using Peroxidase Conjugated Streptavidin and ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1:4,000 to 1:16,000 of the reconstitution concentration is suggested for this product.
• Readout System:
  • Streptavidin Native Protein NBP2-29367
  • Streptavidin Native Protein NBP2-29370
  • Streptavidin Native Protein NBP2-29371

Reactivity Notes

Human, Rabbit and Mouse. Not yet tested in other species.

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
• Preservative: 0.01% Sodium Azide
• Concentration: LYOPH
• Purity: Whole Antiserum
• Reconstitution Instructions: Reconstitute with 100 ul deionized water (or equivalent)

Notes

Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Alternate Names for Aldolase A Antibody [Biotin]

• ALDA
• aldolase A, fructose-bisphosphate
• EC 4.1.2.13
• fructose-1,6-bisphosphate triosephosphate-lyase
• fructose-bisphosphate aldolase A
• GSD12
• Lung cancer antigen NY-LU-1
• MGC10942
• MGC17716
• MGC17767
• Muscle-type aldolase

Background

This gene product, Aldolase A (fructose bisphosphate aldolase) is a glycolytic enzyme that catalyzes the reversible conversion of fructose-1,6-bisphosphate to glyceraldehyde 3 phosphate and dihydroxyacetone phosphate. Three aldolase isozymes (A, B, and C), encoded by three different genes, are differentially expressed during development. Aldolase A is found in the developing embryo and is produced in even greater amounts in adult muscle. Aldolase A expression is repressed in adult liver, kidney and intestine and similar to aldolase C levels in brain and other nervous tissue. Aldolase A deficiency has been associated with myopathy and hemolytic anemia. Alternative splicing of this gene results in multiple transcript variants which encode the same protein.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: ALDOA
• Uniprot:
  • P00883(Rabbit)