Day: December 27, 2024

C-Myc (EQKLISEEDL)-FITC Antibody [Polyclonal]

Sue Steward Main

The C-Myc (EQKLISEEDL)-FITC Antibody is a premium polyclonal antibody conjugated with fluorescein isothiocyanate (FITC), designed to specifically detect the C-Myc epitope tag. Widely used in molecular biology, the C-Myc tag facilitates the detection, purification, and localization of recombinant proteins. The FITC conjugation enables direct fluorescent detection, making it ideal for a variety of applications including flow cytometry, immunofluorescence, and confocal microscopy.

This antibody is optimized for reliable detection of C-Myc-tagged proteins in a range of expression systems and is suitable for high-sensitivity studies in both basic and applied research.

Key Features

  1. Epitope Targeting: Recognizes the highly specific EQKLISEEDL sequence of the C-Myc tag.
  2. Fluorescent Labeling: Conjugated with FITC, a bright and photostable fluorophore with excitation/emission maxima at 495/519 nm, producing sharp green fluorescence.
  3. Polyclonal Design: Detects multiple epitopes, enhancing sensitivity and reducing false negatives.
  4. Versatile Applications: Compatible with immunofluorescence (IF), flow cytometry (FC), confocal microscopy, and immunoprecipitation (IP).
  5. Cross-Species Compatibility: Reacts with C-Myc-tagged proteins across human, mouse, bacterial, and yeast systems.

Specifications

Attribute Details
Host Species Rabbit
Isotype IgG
Conjugate FITC (Fluorescein Isothiocyanate)
Epitope Recognized C-Myc Tag Sequence (EQKLISEEDL)
Concentration 0.1 mg/mL
Applications IF, FC, IP, Confocal Microscopy
Storage Conditions Store at 4°C in the dark
Formulation PBS, pH 7.4, with 0.09% sodium azide and 50% glycerol

Applications

1. Immunofluorescence (IF)

  • Objective: Detect C-Myc-tagged proteins within fixed cells or tissues.
  • Protocol:
    1. Fix samples in 4% paraformaldehyde for 10-15 minutes.
    2. Permeabilize with 0.1% Triton X-100 for 10 minutes.
    3. Block nonspecific binding with 5% BSA or serum for 30 minutes.
    4. Incubate with the C-Myc-FITC Antibody (1:100 dilution) for 1 hour at room temperature.
    5. Wash, counterstain with DAPI for nuclei, and mount for imaging.

2. Flow Cytometry (FC)

  • Objective: Quantify expression of C-Myc-tagged proteins in cell populations.
  • Protocol:
    1. Prepare single-cell suspensions and fix cells using a flow-compatible buffer.
    2. Incubate with the antibody (1:50 dilution) for 30 minutes on ice.
    3. Wash and analyze on a flow cytometer equipped with a 488 nm laser.

3. Confocal Microscopy

  • Objective: High-resolution visualization of C-Myc-tagged protein localization within cells.
  • Protocol:
    • Use the immunofluorescence protocol, and ensure optimal imaging settings for FITC fluorescence.

4. Immunoprecipitation (IP)

  • Objective: Enrich C-Myc-tagged proteins for functional or structural analysis.
  • Protocol:
    1. Add antibody to lysates containing C-Myc-tagged proteins and incubate overnight at 4°C.
    2. Add protein A/G beads to capture the immune complex.
    3. Wash beads to remove nonspecific proteins, then elute bound protein for analysis.

5. Western Blot Validation

  • Objective: Confirm C-Myc expression using fluorescence instead of HRP.
  • Protocol:
    • Run SDS-PAGE and transfer to a PVDF membrane.
    • Use the FITC-antibody directly for fluorescence-based detection.

Research Applications

  1. Protein Expression Studies: Track recombinant protein expression and localization.
  2. Drug Screening: Quantify the effects of small molecules on tagged proteins.
  3. Molecular Interaction Assays: Investigate protein-protein or protein-DNA interactions.
  4. Cell Biology: Visualize and quantify subcellular localization and trafficking of tagged proteins.
  5. High-Throughput Screening: Ideal for assays requiring rapid fluorescence detection.

Advantages

  1. Direct Detection: No need for secondary antibodies, reducing assay time.
  2. Bright Fluorescence: FITC provides excellent signal strength and photostability.
  3. Wide Compatibility: Suitable for use with various recombinant protein systems.
  4. Cost-Effective: Saves time and resources in fluorescent labeling and detection.

Storage and Handling

  • Storage: Store at 4°C in the dark to prevent photobleaching.
  • Stability: Stable for 6 months when stored properly. For long-term use, aliquot and avoid freeze-thaw cycles.

Safety Information

  • Contains sodium azide as a preservative. Use personal protective equipment when handling and dispose of waste properly.

References

  1. PubMed: C-Myc Research Articles
  2. KEGG Pathways: C-Myc Signaling Pathways
  3. NIH: Protein Expression Resources
  4. Addgene: C-Myc Plasmids and Tools