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Product Info Summary
| SKU: | A05022 |
|---|---|
| Size: | 2mL |
| Reactive Species: | Human, Rabbit |
| Host: | Goat |
| Application: | ELISA, IP, WB |
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Product info
Product Name
Anti-Aldolase ALDOA Antibody
SKU/Catalog Number
A05022
Size
2mL
Form
Lyophilized
Description
Boster Bio Anti-Aldolase ALDOA Antibody (Catalog # A05022). Tested in IP, WB applications. This antibody reacts with Human, Rabbit.
Storage & Handling
Store vial at 4°C prior to restoration. For extended storage aliquot contents and freeze at -20°C 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 4°C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening. (Ship at ambient temperature.)
Cite This Product
Anti-Aldolase ALDOA Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05022)
Host
Goat
Contents
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01% (w/v) Sodium Azide
Clonality
Polyclonal
Isotype
Antiserum
Immunogen
Aldolase [Rabbit Muscle]
Reactive Species
A05022 is reactive to ALDOA in Human, Rabbit
Observed Molecular Weight
68 kDa
Background of ALDOA
Aldolase plays a key role in glycolysis and gluconeogenesis. In addition, it may also function as scaffolding protein. In vertebrates, three forms of this ubiquitous glycolytic enzyme are found, aldolase A in muscle, aldolase B in the liver, and aldolase C in the brain. Alkylation of Arg-43 inactivates the enzyme. Aldolase is involved in step 4 of the subpathway that synthesizes D-glyceraldehyde 3-phosphate and glycerone phosphate from D-glucose.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A05022 is guaranteed for ELISA, IP, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
ELISA: 1:5,000 - 1:20,000
IP: 1:100
WB: 1:500 - 1:5,000
Anti-Aldolase has been tested by immunoprecipitation and western blot and is suitable to be assayed against 1.0 µg of Aldolase [Rabbit Muscle] in a standard ELISA using Peroxidase conjugated Affinity Purified anti-Goat IgG [H&L] (Rabbit) code #605-4302 and (ABTS (2,2’-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) code # ABTS-100 as a substrate for 30 minutes at room temperature. A working dilution of 1:3,000 to 1:12,000 of the reconstitution concentration is suggested for this product. Use approximately 5 ul of antibody to immunoprecipitate 50 ul of protein lysate.
Validation Images & Assay Conditions
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Immunoprecipitation of rabbit anti Aldolase antiserum – Immunoprecipitation performed 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 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.
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IgG purified antibody to rabbit muscle aldolase (100-1141, 200-1141 and 200-1341) 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). Other detection methods will yield similar results. This antibody clearly detects a band at ~41 kDa consistent with human aldolase.
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