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Product Info Summary
| SKU: | A01475-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-p70 S6K/RPS6KB1 Antibody Picoband®
SKU/Catalog Number
A01475-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-p70 S6K/RPS6KB1 Antibody Picoband® catalog # A01475-3. Tested in WB,FCM,ELISA applications. This antibody reacts with Human. 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-p70 S6K/RPS6KB1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01475-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Immunogen
E.coli-derived human p70 S6K/RPS6KB1 recombinant protein (Position: R7-Q500). Human RPS6KB1 shares 99.4% and 99.6% amino acid (aa) sequence identity with mouse and rat RPS6KB1, respectively.
Reactive Species
A01475-3 is reactive to RPS6KB1 in Human
Observed Molecular Weight
68 kDa
Calculated molecular weight
59.1 kDa
Background of RPS6KB1
This gene encodes a member of the ribosomal S6 kinase family of serine/threonine kinases. The encoded protein responds to mTOR (mammalian target of rapamycin) signaling to promote protein synthesis, cell growth, and cell proliferation. Activity of this gene has been associated with human cancer. Alternatively spliced transcript variants have been observed. The use of alternative translation start sites results in isoforms with longer or shorter N-termini which may differ in their subcellular localizations. There are two pseudogenes for this gene on chromosome 17.
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
A01475-3 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
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human MCF-7 whole cell, human A549 whole cell, human HepG2 whole cell, human U251 whole cell
FCM: U251 cells
Validation Images & Assay Conditions
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Western blot analysis of P70 S6K/RPS6KB1 using anti-P70 S6K/RPS6KB1 antibody (A01475-3).
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 MCF-7 whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human U251 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-P70 S6K/RPS6KB1 antigen affinity purified polyclonal antibody (Catalog # A01475-3) 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 P70 S6K/RPS6KB1 at approximately 68 kDa. The expected band size for P70 S6K/RPS6KB1 is at 59 kDa.
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Flow Cytometry analysis of U251 cells using anti-P70 S6K/RPS6KB1 antibody (A01475-3).
Overlay histogram showing U251 cells stained with A01475-3 (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-P70 S6K/RPS6KB1 Antibody (A01475-3, 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.
Specific Publications For Anti-p70 S6K/RPS6KB1 Antibody Picoband® (A01475-3)
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