Anti-CBL Antibody Picoband™

Product Name

Anti-CBL Antibody Picoband™

View all CBL Antibodies

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.

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).

*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

A00152-2 is reactive to CBL in Human, Mouse, Rat

Applications

A00152-2 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

120 kDa

Calculated molecular weight

99.633kDa

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.

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Reconsitution

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

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, 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. 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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Figure 2. 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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Figure 3. Flow Cytometry analysis of U87 cells using anti-CBL antibody (A00152-2).
Overlay histogram showing U87 cells stained with A00152-2 (Blue line). 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 (Red line) was also used as a control.

Gene/Protein Information For CBL (Source: Uniprot.org, NCBI)

Alternative Names

Cas-Br-M (murine) ecotropic retroviral transforming sequence; Casitas B-lineage lymphoma proto-oncogene; CBL; CBL2; CBL2E3 ubiquitin-protein ligase CBL; EC 6.3.2; EC 6.3.2.-; oncogene CBL2; Proto-oncogene c-Cbl; RING finger protein 55; RNF55; RNF55C-CBL; Signal transduction protein CBL CBL C-CBL2, FRA11B, NSLL, RNF55, CBL Cbl proto-oncogene E3 ubiquitin-protein ligase CBL|Cas-Br-M (murine) ecotropic retroviral transforming sequence|Cbl proto-oncogene, E3 ubiquitin protein ligase|RING finger protein 55|RING-type E3 ubiquitin transferase CBL|casitas B-lineage lymphoma proto-oncogene|fragile site, folic acid type, rare, fra(11)(q23.3)|oncogene CBL2|proto-oncogene c-Cbl|signal transduction protein CBL

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