Anti-RNF216 Antibody Picoband™

Product Name

Anti-RNF216 Antibody Picoband™

View all TRIAD3 Antibodies

SKU/Catalog Number

A05841-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-RNF216 Antibody Picoband™ catalog # A05841-1. Tested in ELISA, Flow Cytometry, 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-RNF216 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05841-1)

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 RNF216 recombinant protein (Position: K538-R813).

*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

A05841-1 is reactive to RNF216 in Human, Mouse, Rat

Applications

A05841-1 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

50 kDa

Calculated molecular weight

Background of TRIAD3

E3 ubiquitin-protein ligase RNF216 is an enzyme that in humans is encoded by the RNF216 gene. This gene encodes a cytoplasmic protein which specifically colocalizes and interacts with the serine/threonine protein kinase, receptor-interacting protein (RIP). Zinc finger domains of the encoded protein are required for its interaction with RIP and for inhibition of TNF- and IL1-induced NF-kappa B activation pathways. The encoded protein may also function as an E3 ubiquitin-protein ligase which accepts ubiquitin from E2 ubiquitin-conjugating enzymes and transfers it to substrates. Several alternatively spliced transcript variants have been described for this locus but the full-length natures of only some are known.

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
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 RNF216 using anti-RNF216 antibody (A05841-1).
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 U2OS whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: rat testis tissue lysates,
Lane 5: rat heart tissue lysates,
Lane 6: mouse testis tissue lysates,
Lane 7: mouse heart 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-RNF216 antigen affinity purified polyclonal antibody (Catalog # A05841-1) 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 RNF216 at approximately 50 kDa. The expected band size for RNF216 is at 99 kDa.

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Figure 2. Flow Cytometry analysis of HepG2 cells using anti-RNF216 antibody (A05841-1).
Overlay histogram showing HepG2 cells stained with A05841-1 (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-RNF216 Antibody (A05841-1, 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 RNF216 (Source: Uniprot.org, NCBI)

Alternative Names

EC 6.3.2.-; ring finger protein 216Ubiquitin-conjugating enzyme 7-interacting protein 1; Triad domain-containing protein 3; TRIAD3Zinc finger protein inhibiting NF-kappa-B; U7I1; UBCE7IP1ubiquitin conjugating enzyme 7 interacting protein 1; ZINE3 ubiquitin-protein ligase RNF216 RNF216 CAHH, TRIAD3, U7I1, UBCE7IP1, ZIN ring finger protein 216 E3 ubiquitin-protein ligase RNF216|RING-type E3 ubiquitin transferase RNF216|triad domain-containing protein 3|ubiquitin-conjugating enzyme 7-interacting protein 1|zinc finger protein inhibiting NF-kappa-B

For more info on RNF216, check out the RNF216 Infographic

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In this infographic, you will see the following information for RNF216: 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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