Anti-Pirh2/RCHY1 Antibody Picoband™

Product Name

Anti-Pirh2/RCHY1 Antibody Picoband™

View all Pirh2 Antibodies

SKU/Catalog Number

A04533-3

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-Pirh2/RCHY1 Antibody Picoband™ catalog # A04533-3. 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-Pirh2/RCHY1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04533-3)

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 Pirh2/RCHY1 recombinant protein (Position: Q17-Q250).

*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

A04533-3 is reactive to RCHY1 in Human, Mouse, Rat

Reconstitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Observed Molecular Weight

21 kDa

Calculated molecular weight

Background of Pirh2

RING finger and CHY zinc finger domain-containing protein 1 is a protein that in humans is encoded by the RCHY1 gene. The protein encoded by this gene has ubiquitin ligase activity. It mediates E3-dependent ubiquitination and proteasomal degradation of target proteins, including tumor protein 53, histone deacetylase 1, and cyclin-dependent kinase inhibitor 1B, thus regulating their levels and cell cycle progression. Alternatively spliced transcript variants encoding different isoforms have been described for this gene.

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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Applications

A04533-3 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

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

Positive Control

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of Pirh2/RCHY1 using anti-Pirh2/RCHY1 antibody (A04533-3).
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 Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Daudi whole cell lysates,
Lane 4: human 293T whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat PC-12 whole cell lysates,
Lane 7: mouse liver 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-Pirh2/RCHY1 antigen affinity purified polyclonal antibody (Catalog # A04533-3) 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 Pirh2/RCHY1 at approximately 21 kDa. The expected band size for Pirh2/RCHY1 is at 30 kDa.

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Figure 2. Flow Cytometry analysis of HepG2 cells using anti-Pirh2/RCHY1 antibody (A04533-3).
Overlay histogram showing HepG2 cells stained with A04533-3 (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-Pirh2/RCHY1 Antibody (A04533-3, 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 RCHY1 (Source: Uniprot.org, NCBI)

Alternative Names

Androgen receptor N-terminal-interacting protein; androgen-receptor N-terminal-interacting protein; ARNIPRING finger protein 199; CHIMPRNF199DKFZp586C1620; CH-rich interacting match with PLAG1; CH-rich-interacting match with PLAG1; E3 ubiquitin-protein ligase Pirh2; EC 6.3.2.-; hARNIP; hPirh2; PIRH2; PRO1996; ring finger and CHY zinc finger domain containing 1; RING finger and CHY zinc finger domain-containing protein 1; Zinc finger protein 363p53-induced RING-H2 protein; ZNF363 RCHY1 ARNIP, CHIMP, PIRH2, PRO1996, RNF199, ZCHY, ZNF363 ring finger and CHY zinc finger domain containing 1 RING finger and CHY zinc finger domain-containing protein 1|CH-rich interacting match with PLAG1|E3 ubiquitin-protein ligase Pirh2|RING finger protein 199|RING-type E3 ubiquitin transferase RCHY1|androgen-receptor N-terminal-interacting protein|p53-induced protein with a RING-H2 domain|ring finger and CHY zinc finger domain containing 1, E3 ubiquitin protein ligase|zinc finger protein 363|zinc finger, CHY-type

For more info on RCHY1, check out the RCHY1 Infographic

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In this infographic, you will see the following information for RCHY1: 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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