Anti-PIR Antibody Picoband™

Product Name

Anti-PIR Antibody Picoband™

View all Pirin Antibodies

SKU/Catalog Number

A03468-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-PIR Antibody Picoband™ catalog # A03468-1. Tested in ELISA, IF, ICC, WB, Flow Cytometry applications. This antibody reacts with Human, Monkey, 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-PIR Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03468-1)

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 PIR recombinant protein (Position: K5-Q266). Human PIR shares 96.6% and 95.8% amino acid (aa) sequence identity with mouse and rat PIR, 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

A03468-1 is reactive to PIR in Human, Monkey, Mouse, Rat

Applications

A03468-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

36 kDa

Calculated molecular weight

32.113kDa

Background of Pirin

Pirin is a protein that in humans is encoded by the PIR gene. This gene encodes a member of the cupin superfamily. The encoded protein is an Fe(II)-containing nuclear protein expressed in all tissues of the body and concentrated within dot-like subnuclear structures. Interactions with nuclear factor I/CCAAT box transcription factor as well as B cell lymphoma 3-encoded oncoprotein suggest the encoded protein may act as a transcriptional cofactor and be involved in the regulation of DNA transcription and replication. Alternatively spliced transcript variants have been described.

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, 0.1-0.25 µg/ml, Human, Monkey, Mouse, 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 PIR using anti-PIR antibody (A03468-1).
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 HepG2 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: monkey COS-7 whole cell lysates,
Lane 4: rat heart tissue lysates,
Lane 5: rat RH-35 whole cell lysates,
Lane 6: mouse NIH/3T3 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-PIR antigen affinity purified polyclonal antibody (Catalog # A03468-1) at 0.25 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 PIR at approximately 95 kDa. The expected band size for PIR is at 95 kDa.

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Figure 2. IF analysis of PIR using anti-PIR antibody (A03468-1).
PIR was detected in an immunocytochemical section of U2OS 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-PIR Antibody (A03468-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 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 HepG2 cells using anti-PIR antibody (A03468-1).
Overlay histogram showing HepG2 cells stained with A03468-1 (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-PIR Antibody (A03468-1, 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 (Red line) was also used as a control.

Gene/Protein Information For PIR (Source: Uniprot.org, NCBI)

Alternative Names

EC 1.13.11.24; pirin (iron-binding nuclear protein); pirin; Probable quercetin 2,3-dioxygenase PIR; Probable quercetinase PIR pirin pirin|pirin (iron-binding nuclear protein)|probable quercetin 2,3-dioxygenase PIR|probable quercetinase

For more info on PIR, check out the PIR Infographic

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