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Product Info Summary
| SKU: | A00152-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-CBL Antibody Picoband®
SKU/Catalog Number
A00152-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-CBL Antibody Picoband® catalog # A00152-2. Tested in ELISA, Flow Cytometry, IHC, WB 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-CBL Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00152-2)
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 CBL recombinant protein (Position: M470-E766).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00152-2 is reactive to CBL in Human, Mouse, Rat
Observed Molecular Weight
120 kDa
Calculated molecular weight
99.6 kDa
Background of CBL
CBL(Cbl proto-oncogene) is also known as C-CBL, RNF55, CBL2 and E3 ubiquitin protein ligase. CBL is mapped to chromosome 11q23.3-qter by molecular characterization of the breakpoints in 2 somatic cell hybrids. The encoded protein is one of the enzymes required for targeting substrates for degradation by the proteasome. This protein mediates the transfer of ubiquitin from ubiquitin conjugating enzymes(E2) to specific substrates. This protein also contains an N-terminal phosphotyrosine binding domain that allows it to interact with numerous tyrosine-phosphorylated substrates and target them for proteasome degradation. As such it functions as a negative regulator of many signal transduction pathways. This gene has been found to be mutated or translocated in many cancers including acute myeloid leukaemia. Mutations in this gene are also the cause of Noonan syndrome-like disorder.
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
A00152-2 is guaranteed for ELISA, Flow Cytometry, IHC, 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, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human HL-60 whole cell, human K562 whole cell, human A549 whole cell, human Raji whole cell, human CCRF-CEM whole cell, human Daudi whole cell, human MCF-7 whole cell, human Hela whole cell, rat testis tissue, mouse Ana-1 whole cell
IHC: human colorectal cancer tissue
FCM: U87 cell
Validation Images & Assay Conditions
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Western blot analysis of CBL using anti-CBL antibody (A00152-2).
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 HL-60 whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: human CCRF-CEM whole cell lysates,
Lane 6: human Daudi whole cell lysates,
Lane 7: human MCF-7 whole cell lysates,
Lane 8: human Hela whole cell lysates,
Lane 9: rat testis tissue lysates,
Lane 10: mouse Ana-1 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-CBL antigen affinity purified polyclonal antibody (Catalog # A00152-2) 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 CBL at approximately 120 kDa. The expected band size for CBL is at 120 kDa.
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IHC analysis of CBL using anti-CBL antibody (A00152-2).
CBL was detected in a paraffin-embedded section of human colorectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CBL Antibody (A00152-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Flow Cytometry analysis of U87 cells using anti-CBL antibody (A00152-2).
Overlay histogram showing U87 cells stained with A00152-2 (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-CBL Antibody (A00152-2, 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-CBL Antibody Picoband® (A00152-2)
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