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Product Info Summary
| SKU: | A09986-2 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, IP, IF, ICC, WB |
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Product info
Product Name
Anti-LYAR Antibody Picoband®
SKU/Catalog Number
A09986-2
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-LYAR Antibody Picoband® catalog # A09986-2. Tested in WB, ICC/IF, IP, ELISA applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-LYAR Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A09986-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Immunogen
E.coli-derived human LYAR recombinant protein (Position: M1-K379). Human LYAR shares 71.9% and 73.7% amino acid (aa) sequence identity with mouse and rat LYAR, respectively.
Reactive Species
A09986-2 is reactive to LYAR in Human, Mouse, Rat
Observed Molecular Weight
44 kDa
Calculated molecular weight
43.6 kDa
Background of LYAR
Cell growth-regulating nucleolar protein is a protein that in humans is encoded by the LYAR gene (Ly-1 antibody reactive clone). LYAR (Ly-1/CD5 antibody reactive clone) is a 45-50 kDa nucleolar protein that was named for the ability of its antibody to cross-react with Ly-1. Its function is unclear; it is known to associate with MYCN and RRP1B, the latter association giving rise to the suggestion that LYAR is involved with RNA metabolism. Human LYAR is 379 amino acids (aa) in length. It contains two C2H2-type Zn finger regions (aa 6-25 and 33-51) followed by one coiled-coil region (aa 175-219) and an NLS (aa 217-222). There are multiple Zn-binding sites and three utilized phosphorylation sites at Ser244, Ser258 and Ser276. Over aa 288-379, human LYAR shares 75% aa identity with mouse LYAR.
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.
Application & Images
Applications
A09986-2 is guaranteed for ELISA, IP, 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, 0.1-0.25 μg/ml, Human, Mouse, Rat
Immunoprecipitation, 0.5-2 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human A431 whole cell, human K562 whole cell, human HEL whole cell, human U251 whole cell, rat testis tissue, rat RH-35 whole cell, mouse testis tissue, mouse NIH/3T3 whole cell
ICC/IF: HELA cell
IP: K562 cell
Validation Images & Assay Conditions
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Western blot analysis of LYAR using anti-LYAR antibody (A09986-2).
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 A431 whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human HEL whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat testis tissue lysates,
Lane 6: rat RH-35 whole cell lysates,
Lane 7: mouse testis tissue lysates,
Lane 8: 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-LYAR antigen affinity purified polyclonal antibody (Catalog # A09986-2) 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 LYAR at approximately 44 kDa. The expected band size for LYAR is at 44 kDa.
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IF analysis of LYAR using anti-LYAR antibody (A09986-2) and anti-Beta Tubulin antibody (M01857-3).
LYAR was detected in immunocytochemical section of HELA 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-LYAR Antibody (A09986-2) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were 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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Immunoprecipitating (IP) LYAR in K562 whole cell lysate.
Western blot analysis of LYAR using anti-LYAR antibody (A09986-2);
Lane 1: K562 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-LYAR antibody in K562 whole cell lysate;
Lane 3: anti-LYAR antibody (2μg) + K562 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-LYAR antigen affinity purified polyclonal antibody (A09986-2) at a dilution of 0.5 μg/mL and probed with a mouse anti-rabbit IgG-HRP secondary antibody (Catalog # BM2007). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for LYAR at approximately 44 kDa. The expected band size for LYAR is at 44 kDa.
Specific Publications For Anti-LYAR Antibody Picoband® (A09986-2)
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