Aldolase A Antibody - BSA Free

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] - 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] - 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).

Immunoprecipitation: Aldolase A Antibody [NB600-915] - Analysis using the Biotin conjugate of NB600-915. Detection of 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.

Western Blot of Goat anti-Aldolase Antibody. Lane 1: Hela lysate

Western Blot: Aldolase A Antibody [NB600-915] - Twenty one more markedly oxidized proteins identified in 6‐week‐old PLP‐tg66/66 mice in the spinal cord. All non‐mitochondrial oxidized proteins tested by Western blot in the spinal cord are normally expressed. Relative protein level expressed as a percentage of control and in reference to gamma ‐tubulin as a loading marker (n = 6/genotype). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29027761/) licensed under a CC-BY license

• Reactivity: Hu, Mu, Rb
• Applications: WB, ELISA, IP
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: LYOPH

Aldolase A Antibody - BSA Free Summary

• Description: This antibody was prepared from monospecific antiserum by a delipidation, salt fractionation and ion exchange chromatography. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum, purified and partially purified Aldolase [Rabbit Muscle].
  
  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.
• 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 1:7000
  • Immunoprecipitation 1:100
  • Western Blot 1:1000-1:5000
• Application Notes: This product has been tested by ELISA, immunoprecipitation, and western blot. This product is assayed against 1.0 ug of Aldolase [Rabbit Muscle] in a standard ELISA using Peroxidase conjugated Affinity Purified anti-Goat IgG [H&L] (Rabbit) and (ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1:10,000 to 1:40,000 is suggested for this product. Use approximately 5 ul of antibody to immunoprecipitate 50 ul of protein lysate.
• Control:

  Aldolase A Overexpression Lysate

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C. 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
• Preservative: 0.01% Sodium Azide
• Concentration: LYOPH
• Purity: Whole Antiserum
• Reconstitution Instructions: Reconstitute with 100 ul deionized water (or equivalent)

Alternate Names for Aldolase A Antibody - BSA Free

• 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.

Publications for Aldolase A Antibody (NB600-915)(3)

Publications using NB600-915

• Ruiz M, Begou M et al. Oxidative stress and mitochondrial dynamics malfunction are linked in Pelizaeus-Merzbacher disease. Brain Pathol 2017-10-13 [PMID: 29027761] (WB, Mouse)
• Nathalie Launay, Montserrat Ruiz, Laia Grau, Francisco J. Ortega, Ekaterina V. Ilieva, Juan José Martínez, Elena Galea, Isidre Ferrer, Erwin Knecht, Aurora Pujol, Stéphane Fourcade Tauroursodeoxycholic bile acid arrests axonal degeneration by inhibiting the unfolded protein response in X-linked adrenoleukodystrophy Acta Neuropathologica 2016-12-21 [PMID: 28004277]
• Galino J, Ruiz M, Fourcade S, Schluter A et al. Oxidative damage compromises energy metabolism in the axonal degeneration mouse model of X-adrenoleukodystrophy. Antioxid Redox Signal 2011-10-15 [PMID: 21453200] (WB, Mouse)

FAQs for Aldolase A Antibody (NB600-915). (Showing 1 - 1 of 1 FAQs).

1. We have to store reference NB600-915 over the weekend and I was wondering if the storage conditions (Store at 4C short term), do apply for such a time or would we be better off putting the antibody to -20C?.
   • The antibody can be stored at 4C over the weekend as it contains sodium azide which is a stabilizer. We recommend -20C for long term storage.

Bioinformatics

• Gene Symbol: ALDOA
• Entrez:
  • 226(Human)
• Uniprot:
  • P00883(Rabbit)