Skip to main content

Anti-EGFP antibody [F56-6A1.2.3] ab184601 is a mouse monoclonal antibody that is used in EGFP western blotting and immunofluorescence.


Images

Immunocytochemistry/ Immunofluorescence - Anti-EGFP antibody [F56-6A1.2.3] (AB184601), expandable thumbnail
  • Western blot - Anti-EGFP antibody [F56-6A1.2.3] (AB184601), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-EGFP antibody [F56-6A1.2.3] (AB184601), expandable thumbnail
  • Western blot - Anti-EGFP antibody [F56-6A1.2.3] (AB184601), expandable thumbnail

Publications

Key facts

Isotype

IgG2b

Host species

Mouse

Storage buffer

Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA

Form

Liquid

Clonality

Monoclonal

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IF
Transfected cell line - Aequorea victoria
Not recommended
Tested
Transfected cell lysate - Aequorea victoria
Tested
Not recommended

Tested
Tested

Species

Transfected cell lysate - Aequorea victoria

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Transfected cell line - Aequorea victoria

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell line - Aequorea victoria

Dilution info

1/20.00000 - 1/100.00000

Notes

-

Not recommended
Not recommended

Species

Transfected cell lysate - Aequorea victoria

Dilution info

-

Notes

-

Target data

Function

Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin.

Alternative names

Green fluorescent protein, GFP

Recommended products

Anti-EGFP antibody [F56-6A1.2.3] ab184601 is a mouse monoclonal antibody that is used in EGFP western blotting and immunofluorescence.

Key facts

Isotype

IgG2b

Form

Liquid

Clonality

Monoclonal

Clone number

F56-6A1.2.3

Purification technique

Affinity purification Protein A

Concentration
Loading...
Purification notes

in vitro produced

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.

Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

EGFP or Enhanced Green Fluorescent Protein is a widely used marker in molecular biology. Known also as GFP EGFP its expression occurs in various organisms and facilitates easy visualization of cellular and molecular processes. The molecular weight of EGFP is approximately 27 kDa contributing to its manageable size for integration into other proteins. Generally scientists observe EGFP in tissues and cell types where it is used as a fusion tag to monitor protein localization and dynamics.

Biological function summary

EGFP functions without disrupting the normal cellular environment where it fluoresces green under ultraviolet or blue light. The protein does not form a complex with other molecules allowing it to remain stable and functional in diverse conditions. Scientists often utilize the EGFP sequence to create fusion proteins aiding in the study of protein-protein interactions trafficking and other cellular processes. This versatility makes EGFP a valuable tool in biological experiments.

Pathways

Scientists exploit EGFP as a marker within research on cellular signaling and gene expression pathways. It is particularly useful within the analysis of the Ras signaling pathway and the NF-kB pathway where it can help track the movement and interaction of key proteins such as Ras and IκB. The fluorescent characteristics of EGFP allow real-time observation of these pathways enhancing the understanding of molecular processes in live cells.

Associated diseases and disorders

EGFP serves as a critical tool in cancer research and neurodegenerative diseases. Through its fusion with proteins like p53 a tumor suppressor researchers can track gene expression and protein stability in cancerous cells. Similarly by merging EGFP with proteins such as tau linked to Alzheimer's disease scientists investigate protein aggregation and neuron behavior in neurodegenerative conditions. Therefore EGFP remains integral to disease research offering insights into the underlying molecular mechanisms.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFP antibody [F56-6A1.2.3] (ab184601), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-EGFP antibody [F56-6A1.2.3] (ab184601)




    Immunofluorescent analysis of GFP Tag was performed using H3-GFP construct transfected in HEK-293E cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ab184601 at 1/100 dilution and Histone H3 Rabbit Polyclonal Antibody at 0.5 μg/mL in 0.1% BSA, incubated at 4°C overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor® Plus 555 and Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor® Plus 647 respectively at a dilution of 1/2000 for 45 minutes at room temperature.

    Image a: Anti-EGFP antibody [F56-6A1.2.3], ab184601 (green)
    Image b: Histone H3 (red)
    Image c: Nuclei stained with DAPI (blue)
    Image d: Merged image showing the co-localization of nuclear signals in transfected cells
    Image e: Untransfected HEK cells
    Image f: Control cells with no primary antibody to assess background

    The images were captured at 60X magnification.


  • Western blot - Anti-EGFP antibody [F56-6A1.2.3] (ab184601), expandable thumbnail

    Western blot - Anti-EGFP antibody [F56-6A1.2.3] (ab184601)

    Western Blot was performed using ab184601 by loading whole cell extracts of untransfected and transiently transfected HEK-293E. Lysates were electrophoresed using a 4-12% Bis-Tris Protein Gel. Resolved proteins were then transferred onto a nitrocellulose membrane using a dry blotting system. A ~45 kDa band of H3-GFP and a ~92 kDa band of p65-GFP were observed in transfected lysates. A 53 kDa recombinant protein consisting multiple epitope tags was used as a positive control for GFP detection. This product also detects Yellow Fluorescent Protein (YFP), a variant of GFP as observed in Lane 7. No cross-reactivity was seen with mCherry (RFP family) expressing lysate.

    All lanes: Western blot - Anti-EGFP antibody [F56-6A1.2.3] (ab184601) at 1/1000 dilution

    Lane 1: Untransfected HEK-293E (human epithelial cell line from embryonic kidney) whole cell lysate at 40 µg

    Lane 2: Empty vector control at 40 µg

    Lane 3: H3-GFP at 40 µg

    Lane 4: H3-GFP at 20 µg

    Lane 5: H3-GFP at 10 µg

    Lane 6: p65-GFP at 40 µg

    Lane 7: p65-YFP at 40 µg

    Lane 8: H3-mCherry at 40 µg

    Lane 9: 53 kDa recombinant protein consisting multiple epitope tags at 0.025 µg

    Secondary

    All lanes: Goat anti-Mouse IgG (H+L) (HRP) at 1/4000 dilution

    Developed using the ECL technique.

    Predicted band size: 27 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFP antibody [F56-6A1.2.3] (ab184601), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-EGFP antibody [F56-6A1.2.3] (ab184601)

    Immunofluorescent analysis of U-2 OS (Human bone osteosarcoma epithelial cell line) cells transfected with an EGFR-EGFP fusion protein (formalin-fixed, 0.1% Triton X-100 permeabilized) using either natural fluorescence (green) or an anti-EGFP antibody (red). Cells were then labeled with ab184601 at 1/20 dilution followed with DyLight 550 goat anti-mouse IgG secondary antibody at 1/200 dilution. Nuclei (blue) were stained with Hoechst 33342 dye. 20X magnification.

  • Western blot - Anti-EGFP antibody [F56-6A1.2.3] (ab184601), expandable thumbnail

    Western blot - Anti-EGFP antibody [F56-6A1.2.3] (ab184601)

    All lanes: Western blot - Anti-EGFP antibody [F56-6A1.2.3] (ab184601) at 1/1000 dilution

    Lane 1: Recombinant GFP at 1 µg

    Lane 2: Recombinant GFP at 0.5 µg

    Lane 3: Recombinant GFP at 0.25 µg

    Lane 4: Recombinant GFP at 0.125 µg

    Lane 5: Recombinant GFP at 0.0625 µg

    Secondary

    All lanes: goat anti-mouse IgG-HRP at 1/15000 dilution

    Developed using the ECL technique.

    Predicted band size: 27 kDa

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com