Anti-NHP2 Antibody Picoband™

Product Name

Anti-NHP2 Antibody Picoband™

View all Nhp2 Antibodies

SKU/Catalog Number

A05665-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-NHP2 Antibody Picoband™ catalog # A05665-1. Tested in ELISA, IF, ICC, WB, Flow Cytometry applications. This antibody reacts with Human.

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-NHP2 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05665-1)

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 NHP2 recombinant protein (Position: E21-Q136).

*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

A05665-1 is reactive to Nhp2 in Human

Reconstitution

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

Observed Molecular Weight

20 kDa

Calculated molecular weight

101272 MW

Background of Nhp2

This gene is a member of the H/ACA snoRNPs (small nucleolar ribonucleoproteins) gene family. snoRNPs are involved in various aspects of rRNA processing and modification and have been classified into two families: C/D and H/ACA. The H/ACA snoRNPs also include the DKC1, NOLA1 and NOLA3 proteins. These four H/ACA snoRNP proteins localize to the dense fibrillar components of nucleoli and to coiled (Cajal) bodies in the nucleus. Both 18S rRNA production and rRNA pseudouridylation are impaired if any one of the four proteins is depleted. The four H/ACA snoRNP proteins are also components of the telomerase complex. This gene encodes a protein related to Saccharomyces cerevisiae Nhp2p. Alternative splicing results in multiple transcript variants.

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

A05665-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, 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
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Positive Control

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of NHP2 using anti-NHP2 antibody (A05665-1).
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 SH-SY5Y whole cell lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: human Hela 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-NHP2 antigen affinity purified polyclonal antibody (Catalog # A05665-1) 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-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 NHP2 at approximately 20 kDa. The expected band size for NHP2 is at 20 kDa.

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Figure 2. IF analysis of NHP2 using anti-NHP2 antibody (A05665-1) and anti-Beta Tubulin antibody (M01857-3).
NHP2 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-NHP2 Antibody (A05665-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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 SH-SY5Y cells using anti-NHP2 antibody (A05665-1).
Overlay histogram showing SH-SY5Y cells stained with A05665-1 (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-NHP2 Antibody (A05665-1, 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 Nhp2 (Source: Uniprot.org, NCBI)

Alternative Names

FLJ20479; H/ACA ribonucleoprotein complex subunit 2; NHP2 ribonucleoprotein homolog (yeast); NHP2-like protein; NHP2P; NOLA2member 2 (H/ACA small nucleolar RNPs); Nucleolar protein family A member 2; snoRNP protein NHP2

For more info on Nhp2, check out the Nhp2 Infographic

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In this infographic, you will see the following information for Nhp2: 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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