Anti-NIRF Antibody Picoband™ (monoclonal, 6B5)

Product Name

Anti-NIRF Antibody Picoband™ (monoclonal, 6B5)

View all NIRF Antibodies

SKU/Catalog Number

M06294-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-NIRF Antibody Picoband™ (monoclonal, 6B5) catalog # M06294-1. Tested in Flow Cytometry, IF, ICC, WB 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-NIRF Antibody Picoband™ (monoclonal, 6B5) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M06294-1)

Host

Mouse

Contents

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

Clonality

Monoclonal

Clone Number

6B5

Isotype

Mouse IgG2b

Immunogen

A synthetic peptide corresponding to a sequence at the N-terminus of human NIRF, identical to the related mouse and rat sequences.

*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

M06294-1 is reactive to Uhrf2 in Human, Rat

Applications

M06294-1 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

90 kDa

Calculated molecular weight

Background of NIRF

E3 ubiquitin-protein ligase UHRF2 is an enzyme that in humans is encoded by the UHRF2 gene. This gene encodes a nuclear protein which is involved in cell-cycle regulation. The encoded protein is a ubiquitin-ligase capable of ubiquinating PCNP (PEST-containing nuclear protein), and together they may play a role in tumorigenesis. The encoded protein contains an NIRF_N domain, a PHD finger, a set- and ring-associated (SRA) domain, and a RING finger domain and several of these domains have been shown to be essential for the regulation of cell proliferation. This protein may also have a role in intranuclear degradation of polyglutamine aggregates. Alternative splicing results in multiple transcript variants some of which are non-protein coding.

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
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x10⁶ cells, Human, Rat

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of NIRF using anti-NIRF antibody (M06294-1).
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 HT1080 whole cell lysates,
Lane 3: human Jurkat 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 mouse anti-NIRF antigen affinity purified monoclonal antibody (Catalog # M06294-1) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for NIRF at approximately 90 kDa. The expected band size for NIRF is at 90 kDa.

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Figure 2. IF analysis of NIRF using anti-NIRF antibody (M06294-1).
NIRF was detected in an immunocytochemical section of Hela 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 mouse anti-NIRF Antibody (M06294-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 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 Hela cells using anti-NIRF antibody (M06294-1).
Overlay histogram showing Hela cells stained with M06294-1 (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 mouse anti-NIRF Antibody (M06294-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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.

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Figure 4. Flow Cytometry analysis of RH35 cells using anti-NIRF antibody (M06294-1).
Overlay histogram showing RH35 cells stained with M06294-1 (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 mouse anti-NIRF Antibody (M06294-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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 Uhrf2 (Source: Uniprot.org, NCBI)

Alternative Names

DKFZp434B0920; DKFZp686G0837; E3 ubiquitin-protein ligase UHRF2; EC 6.3.2; EC 6.3.2.-; MGC33463; NIRFUbiquitin-like-containing PHD and RING finger domains protein 2; Np95/ICBP90-like RING finger protein; Np95-like RING finger protein; Nuclear protein 97; Nuclear zinc finger protein Np97; RNF107RING finger protein 107; Ubiquitin-like PHD and RING finger domain-containing protein 2; ubiquitin-like with PHD and ring finger domains 2; ubiquitin-like, containing PHD and RING finger domains, 2; URF2 Uhrf2|2310065A22Rik, AI426270, AW214556, D130071B19Rik, N, Nirf|ubiquitin-like, containing PHD and RING finger domains 2|E3 ubiquitin-protein ligase UHRF2|Np95-like ring finger protein|RING-type E3 ubiquitin transferase UHRF2|nuclear protein 97|nuclear zinc finger protein Np97|ubiquitin-like PHD and RING finger domain-containing protein 2|ubiquitin-like-containing PHD and RING finger domains protein 2

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In this infographic, you will see the following information for Uhrf2: 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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