RNF216 Recombinant antibody

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  WB analysis of SW480 using 83658-2-RR

  SW480 cells were subjected to SDS PAGE followed by western blot with 83658-2-RR (RNF216 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of Caco-2 using 83658-2-RR

  Caco-2 cells were subjected to SDS PAGE followed by western blot with 83658-2-RR (RNF216 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of SW480 using 83658-2-RR

  IP result of anti-RNF216 (IP:83658-2-RR, 4ug; Detection:83658-2-RR 1:1000) with SW480 cells lysate 1280 ug.

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  FC experiment of A431 using 83658-2-RR

  1x10^6 A431 cells were intracellularly stained with 0.25 ug RNF216 Recombinant antibody (83658-2-RR, Clone:240489G2) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 83658-2-RR

  Biolayer interferometry (BLl) kinetic assays of 83658-2-RR against Human RNF216 were performed. The affinity constant is below 1 pM.

RNF216 Recombinant Antibody for WB, FC (Intra), IP, ELISA

Cat No. 83658-2-RR

Clone No.240489G2

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, FC (Intra), IP, ELISA

Synonyms

Triad domain-containing protein 3, RING-type E3 ubiquitin transferase RNF216, RING finger protein 216, EC:2.3.2.27, E3 ubiquitin-protein ligase RNF216

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Tested Applications

• Positive WB detected in: SW480 cells, Caco-2 cells
• Positive IP detected in: SW480 cells
• Positive FC (Intra) detected in: A431 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

83658-2-RR targets RNF216 in WB, FC (Intra), IP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: RNF216 fusion protein Ag0238
• Full Name: ring finger protein 216
• Calculated Molecular Weight: 48 kDa
• Observed Molecular Weight: ~130 kDa
• GenBank Accession Number: BC000787
• Gene Symbol: RNF216
• Gene ID (NCBI): 54476
• RRID: AB_3671266
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: Q9NWF9
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

RNF216/TRIAD3 is an RBR (RING-between-RING) E3 ligase that encodes for multiple isoforms that include TRIAD3A, TRIAD3B, TRIAD3C and TRIAD3D/E. RNF216 is a broadly expressed RBR E3 ligase, and loss-of-function mutations in RNF216 are linked to the neurode-generative conditions Gordon-Holmes syndrome (GHS) and Huntington-like disorders (PMID: 34998453, 35620441). RNF216 has three different molecular weights, 99 kDa, 106 kDa and 57 kDa, respectively. Western analysis detected the ~130 kDa band as indicated in literature (PMID: 21270397).