Anti-FBXO32 Antibody Picoband™

Product Name

Anti-FBXO32 Antibody Picoband™

View all Fbx32 Antibodies

SKU/Catalog Number

A02531-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-FBXO32 Antibody Picoband™ catalog # A02531-1. Tested in ELISA, IF, ICC, WB, Flow Cytometry applications. This antibody reacts with Human, Mouse.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-FBXO32 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02531-1)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

IgG

Immunogen

E.coli-derived human FBXO32 recombinant protein (Position: Q6-F355).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross reactivity with other proteins.

Reactive Species

A02531-1 is reactive to Fbxo32 in Human, Mouse

Applications

A02531-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

42 kDa

Calculated molecular weight

52588 MW

Background of Fbx32

F-box only protein 32, also known as "MAFbx", for "Muscle Atrophy F-box gene", and "Atrogin-1," is a protein that in humans is encoded by the FBXO32 gene. This gene encodes a member of the F-box protein family which is characterized by an approximately 40 amino acid motif, the F-box. The F-box proteins constitute one of the four subunits of the ubiquitin protein ligase complex called SCFs (SKP1-cullin-F-box), which function in phosphorylation-dependent ubiquitination. The F-box proteins are divided into 3 classes: Fbws containing WD-40 domains, Fbls containing leucine-rich repeats, and Fbxs containing either different protein-protein interaction modules or no recognizable motifs. The protein encoded by this gene belongs to the Fbxs class and contains an F-box domain. This protein is highly expressed during muscle atrophy, whereas mice deficient in this gene were found to be resistant to atrophy. This protein is thus a potential drug target for the treatment of muscle atrophy. Alternative splicing results in multiple transcript variants encoding different isoforms.

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.

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Reconstitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

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.

Western blot, 0.25-0.5 µg/ml, Mouse
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Fbx32/FBXO32 using anti-Fbx32/FBXO32 antibody (A02531-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: mouse heart tissue lysates,
Lane 2: mouse skeletal muscle tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Fbx32/FBXO32 antigen affinity purified polyclonal antibody (Catalog # A02531-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Fbx32/FBXO32 at approximately 42 kDa. The expected band size for Fbx32/FBXO32 is at 42 kDa.

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Figure 2. IF analysis of Fbx32/FBXO32 using anti-Fbx32/FBXO32 antibody (A02531-1) and anti-Beta Tubulin antibody (M01857-3).
Fbx32/FBXO32 was detected in immunocytochemical section of HELA cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-Fbx32/FBXO32 Antibody (A02531-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of PC-3 cells using anti-Fbx32/FBXO32 antibody (A02531-1).
Overlay histogram showing PC-3 cells stained with A02531-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Fbx32/FBXO32 Antibody (A02531-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For Fbxo32 (Source: Uniprot.org, NCBI)

Alternative Names

atrogin 1; ATROGIN1; atrogin-1; F-box only protein 32; F-box protein 32; Fbx32; FLJ32424; MAFBX; MAFbxMGC33610; Muscle atrophy F-box protein Fbxo32|4833442G10Rik, AI430017, ATR, ATROGIN1, Gm20361, MAFb, MAFbx, atro|F-box protein 32|F-box only protein 32|atrogin 1|muscle atrophy F-box protein

For more info on Fbxo32, check out the Fbxo32 Infographic

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In this infographic, you will see the following information for Fbxo32: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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