Anti-USP5 Antibody Picoband™

Product Name

Anti-USP5 Antibody Picoband™

View all Isopeptidase T/USP5 Antibodies

SKU/Catalog Number

A04550-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-USP5 Antibody Picoband™ catalog # A04550-1. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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-USP5 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04550-1)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human USP5 recombinant protein (Position: M1-K800).

*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

A04550-1 is reactive to USP5 in Human, Mouse, Rat

Applications

A04550-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

96 kDa

Calculated molecular weight

Background of Isopeptidase T/USP5

Ubiquitin specific peptidase 5 is an enzyme that in humans is encoded by the USP5 gene. Ubiquitin dependent proteolysis is a complex pathway of protein metabolism implicated in such diverse cellular functions as maintenance of chromatin structure, receptor function, and degradation of abnormal proteins. A late step of the process involves disassembly of the polyubiquitin chains on degraded proteins into ubiquitin monomers. USP5 disassembles branched polyubiquitin chains by a sequential exo mechanism, starting at the proximal end of the chain.

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

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of USP5 using anti-USP5 antibody (A04550-1).
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: human A549 whole cell lysates,
Lane 2: human U20S whole cell lysates,
Lane 3: human MDA-MB-453 whole cell lysates,
Lane 4: huamn MCF-7 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat testis tissue lysates,
Lane 7: mouse brain 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-USP5 antigen affinity purified polyclonal antibody (Catalog # A04550-1) at 0.25 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 USP5 at approximately 96 kDa. The expected band size for USP5 is at 96 kDa.

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Figure 2. IF analysis of USP5 using anti-USP5 antibody (A04550-1).
USP5 was detected in an immunocytochemical section of A549 cells. 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-USP5 Antibody (A04550-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of U251 cells using anti-USP5 antibody (A04550-1).
Overlay histogram showing U251 cells stained with A04550-1 (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-USP5 Antibody (A04550-1, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Gene/Protein Information For USP5 (Source: Uniprot.org, NCBI)

Alternative Names

Deubiquitinating enzyme 5; EC 3.1.2.15; EC 3.4.19.12; Isopeptidase T; IsoT; ISOTubiquitin carboxyl-terminal hydrolase 5; ubiquitin isopeptidase T; ubiquitin specific peptidase 5 (isopeptidase T); ubiquitin specific protease 5 (isopeptidase T); ubiquitin thioesterase 5; Ubiquitin thiolesterase 5; ubiquitin-specific protease-5 (ubiquitin isopeptidase T); Ubiquitin-specific-processing protease 5; USP5 USP5 ISOT ubiquitin specific peptidase 5 ubiquitin carboxyl-terminal hydrolase 5|deubiquitinating enzyme 5|isopeptidase T|testicular tissue protein Li 218|ubiquitin isopeptidase T|ubiquitin specific protease 5 (isopeptidase T)|ubiquitin thioesterase 5|ubiquitin thiolesterase 5|ubiquitin-specific protease-5 (ubiquitin isopeptidase T)|ubiquitin-specific-processing protease 5

For more info on USP5, check out the USP5 Infographic

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In this infographic, you will see the following information for USP5: 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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