CBL ELISA Kit

Catalog No. ABIN8068181

Human CBL ELISA Kit, Colorimetric assay for quantification of Human CBL.

Quick Overview for CBL ELISA Kit (ABIN8068181)

Target: CBL (Cas-Br-M (Murine) Ecotropic Retroviral Transforming Sequence (CBL))

Binding Specificity: total, ubiquitinated

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details CBL ELISA Kit

Purpose: Measures ubiquitinated CBL and total CBL in human cell and tissue lysates, enabling precise assessment of CBL ubiquitination by ELISA. This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com.

Analytical Method: Quantitative

Components:

Size / Qty
• Microplate Coated With Anti-Pan-CBL Antibody
• Anti-Ubiquitination Detection Antibody
• Anti-CBL Detection Antibody
• HRP-Conjugated Streptavidin
• 200 μL
• Wash Buffer
• Assay Diluent
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Microplate reader capable of measuring absorbance at 450 nm
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Log-log graph paper or computer and software for ELISA data analysis
• Absorbent paper
• Tubes to prepare sample dilutions
• Distilled or deionized water
• Shaker

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed.
2. Add 100 μL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
3. Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
4. Add 100 μL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

Target Details for CBL

Target: CBL (Cas-Br-M (Murine) Ecotropic Retroviral Transforming Sequence (CBL))

Alternative Name: CBL

Background:

Gene Symbols: CBL, CBL2, RNF55

Background: The Ubiquitinated-CBL and Total-CBL ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human ubiquitinated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan CBL antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and CBL present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-ubiquitination antibody is used to detect ubiquitinated CBL or biotinylated anti-CBL antibody is used to detect total CBL . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ubiquitinated CBL or total CBL bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): E3 ubiquitin-protein ligase CBL, Casitas B-lineage lymphoma proto-oncogene, Proto-oncogene c-Cbl, RING finger protein 55, RING-type E3 ubiquitin transferase CBL, Signal transduction protein CBL

Gene ID: 867

UniProt: P22681

Pathways: TCR Signaling, Interferon-gamma Pathway, EGFR Signaling Pathway, EGFR Downregulation, VEGFR1 Specific Signals