Anti-BRCC36/BRCC3 Antibody Picoband™

Product Name

Anti-BRCC36/BRCC3 Antibody Picoband™

View all BRCC3 Antibodies

SKU/Catalog Number

A04690-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-BRCC36/BRCC3 Antibody Picoband™ catalog # A04690-1. 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-BRCC36/BRCC3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04690-1)

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 BRCC36/BRCC3 recombinant protein (Position: Q10-M276).

*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

A04690-1 is reactive to BRCC3 in Human, Mouse, Rat

Applications

A04690-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

36 kDa

Calculated molecular weight

Background of BRCC3

Lys-63-specific deubiquitinase BRCC36 is an enzyme that in humans is encoded by the BRCC3 gene. This gene encodes a subunit of the BRCA1-BRCA2-containing complex (BRCC), which is an E3 ubiquitin ligase. This complex plays a role in the DNA damage response, where it is responsible for the stable accumulation of BRCA1 at DNA break sites. The component encoded by this gene can specifically cleave Lys 63-linked polyubiquitin chains, and it regulates the abundance of these polyubiquitin chains in chromatin. The loss of this gene results in abnormal angiogenesis and is associated with syndromic moyamoya, a cerebrovascular angiopathy. Alternative splicing results in multiple transcript variants. A related pseudogene has been identified on chromosome 5.

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, 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 BRCC36/BRCC3 using anti-BRCC36/BRCC3 antibody (A04690-1).
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 HEK293 whole cell lysates,
Lane 2: rat kidney tissue lysates,
Lane 3: rat pancreas tissue lysates,
Lane 4: mouse kidney 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-BRCC36/BRCC3 antigen affinity purified polyclonal antibody (Catalog # A04690-1) 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 BRCC36/BRCC3 at approximately 36 kDa. The expected band size for BRCC36/BRCC3 is at 36 kDa.

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Figure 2. IF analysis of BRCC36/BRCC3 using anti-BRCC36/BRCC3 antibody (A04690-1).
BRCC36/BRCC3 was detected in an immunocytochemical section of A431 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-BRCC36/BRCC3 Antibody (A04690-1) 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 U87 cells using anti-BRCC36/BRCC3 antibody (A04690-1).
Overlay histogram showing U87 cells stained with A04690-1 (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-BRCC36/BRCC3 Antibody (A04690-1, 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 BRCC3 (Source: Uniprot.org, NCBI)

Alternative Names

BRCA1/BRCA2-containing complex subunit 3; BRCA1/BRCA2-containing complex subunit 36; BRCA1/BRCA2-containing complex, subunit 3; BRCA1-A complex subunit BRCC36; BRCC36C6.1ACXorf53; BRISC complex subunit BRCC36; c6.1A; chromosome X open reading frame 53; EC 3.1.2.15; EC 3.4.19.-; lys-63-specific deubiquitinase BRCC36 BRCC3 BRCC36, C6.1A, CXorf53 BRCA1/BRCA2-containing complex subunit 3 lys-63-specific deubiquitinase BRCC36|BRCA1-A complex subunit BRCC36|BRCA1/BRCA2-containing complex subunit 36|BRISC complex subunit BRCC36

For more info on BRCC3, check out the BRCC3 Infographic

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In this infographic, you will see the following information for BRCC3: 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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