Anti-RING1 Antibody Picoband™

Product Name

Anti-RING1 Antibody Picoband™

View all RING1 Antibodies

SKU/Catalog Number

A01824-2

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-RING1 Antibody Picoband™ catalog # A01824-2. Tested in WB, ICC/IF, FCM, ELISA applications. This antibody reacts with Human, 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-RING1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01824-2)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Clone Number

1B9

Isotype

IgG

Immunogen

E.coli-derived human RING1 recombinant protein (Position: N6-K387). Human RING1 shares 96.3% and 96.6% amino acid (aa) sequence identity with mouse and rat RING1, respectively.

*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

A01824-2 is reactive to RING1 in Human, Rat

Applications

A01824-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

50 kDa

Calculated molecular weight

42.429kDa

Background of RING1

E3 ubiquitin-protein ligase RING1 is an enzyme that in humans is encoded by the RING1 gene. This gene belongs to the RING finger family, members of which encode proteins characterized by a RING domain, a zinc-binding motif related to the zinc finger domain. The gene product can bind DNA and can act as a transcriptional repressor. It is associated with the multimeric polycomb group protein complex. The gene product interacts with the polycomb group proteins BMI1, EDR1, and CBX4, and colocalizes with these proteins in large nuclear domains. It interacts with the CBX4 protein via its glycine-rich C-terminal domain. The gene maps to the HLA class II region, where it is contiguous with the RING finger genes FABGL and HKE4.

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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Reconsitution

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, 0.5-1 µg/ml, Human, Rat
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 RING1 using anti-RING1 antibody (A01824-2).
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 HEL whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: rat PC-12 whole cell 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-RING1 antigen affinity purified polyclonal antibody (Catalog # A01824-2) 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 RING1 at approximately 50 kDa. The expected band size for RING1 is at 43 kDa.

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Figure 2. IF analysis of RING1 using anti-RING1 antibody (A01824-2) and anti-Beta Tubulin antibody (M01857-3).
RING1 was detected in immunocytochemical section of A549 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-RING1 Antibody (A01824-2) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and FITC Conjugated Goat Anti-Mouse IgG (BA1101) 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 JK cells using anti-RING1 antibody (A01824-2).
Overlay histogram showing JK cells stained with A01824-2 (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-RING1 Antibody (A01824-2, 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 RING1 (Source: Uniprot.org, NCBI)

Alternative Names

EC 6.3.2; EC 6.3.2.-; Polycomb complex protein RING1; Really interesting new gene 1 protein; ring finger protein 1RING1A; RNF1E3 ubiquitin-protein ligase RING1 RING1 RING1A, RNF1 ring finger protein 1 E3 ubiquitin-protein ligase RING1|RING-type E3 ubiquitin transferase RING1|polycomb complex protein RING1|really interesting new gene 1 protein

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