Product Info Summary
| SKU: | A06618-1 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-PIK3R4 Antibody Picoband®
SKU/Catalog Number
A06618-1
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-PIK3R4 Antibody Picoband® catalog # A06618-1. Tested in ELISA, WB, Flow Cytometry 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-PIK3R4 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A06618-1)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
IgG
Immunogen
E.coli-derived human PIK3R4 recombinant protein (Position: R36-Q1339). Human PIK3R4 shares 95.7% and 95.6% amino acid (aa) sequence identity with mouse and rat PIK3R4, respectively.
Cross-reactivity
No cross reactivity with other proteins.
Reactive Species
A06618-1 is reactive to PIK3R4 in Human
Observed Molecular Weight
150 kDa
Calculated molecular weight
153.1 kDa
Background of PIK3R4
Phosphoinositide 3-kinase regulatory subunit 4, also known as PI3-kinase regulatory subunit 4 or PI3-kinase p150 subunit or phosphoinositide 3-kinase adaptor protein, or VPS15 is an enzyme that in humans is encoded by the PIK3R4 gene. The serine/threonine PI3 kinase regulatory subunit 4 (PIK3R4, Vps15) is the mammalian homologue of the yeast vacuolar protein sorting 15. PIK3R4 regulates the kinase activity of PI3K class III and anchors the kinase to cellular membranes through myristoylation. Recruitment of PI3K class III to the site of early endosome fusion and docking is directly mediated by PIK3R4 binding to the small GTPase Rab5 through its HEAT and WD-40 domains. The PIK3R4/PI3K class III plays a role in late endosome function through PIK3R4 binding to the Rab7 GTPase. In addition to its role in trafficking, the PIK3R4/PI3K class III complex interacts with beclin-1 to play a role during several stages of autophagy. Autophagosome formation is stimulated when Atg14 complexes with PIK3R4, PI3K class III, and beclin-1. The UVRAG protein competes with Atg14 for beclin-1 binding, forming a mutually exclusive complex with PIK3R4, PI3K class III, and beclin-1 that regulates autophagosome maturation. Autophagosome maturation is impaired in the presence of the beclin-1-binding protein Rubicon. Co-expression of PIK3R4 is required for PI3K class III activation and regulation by both beclin-1/UVRAG and by nutrients. Overexpression of PIK3R4 protein has been associated with decreased survival in patients with ovarian tumors, while mutations of the corresponding PIK3R4 gene are associated with metastatic melanoma, suggesting that PIK3R4 functions in cancer.
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
A06618-1 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 Hela whole cell, human HepG2 whole cell, human 293T whole cell
FCM: HepG2 cell
Validation Images & Assay Conditions
Click image to see more details
Western blot analysis of VPS15/PIK3R4 using anti-VPS15/PIK3R4 antibody (A06618-1).
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 Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human 293T 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-VPS15/PIK3R4 antigen affinity purified polyclonal antibody (Catalog # A06618-1) 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 VPS15/PIK3R4 at approximately 150 kDa. The expected band size for VPS15/PIK3R4 is at 153 kDa.
Click image to see more details
Flow Cytometry analysis of HepG2 cells using anti-VPS15/PIK3R4 antibody (A06618-1).
Overlay histogram showing HepG2 cells stained with A06618-1 (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-VPS15/PIK3R4 Antibody (A06618-1, 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.
Specific Publications For Anti-PIK3R4 Antibody Picoband® (A06618-1)
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