• Product name
    Anti-GFP antibody [6AT316]
    See all GFP primary antibodies
  • Description
    Mouse monoclonal [6AT316] to GFP
  • Host species
  • Specificity
    Detects both GFP and YFP by Western Blotting.
  • Tested applications
    Suitable for: IHC-Fr, ELISA, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Full length native protein (purified) corresponding to GFP.
    Database link: P42212


  • Form
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer
    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein G purified
  • Purification notes
    This antibody was purified by protein G and Ni-NTA nickel affinity chromatography, followed by dialysis against PBS.
  • Clonality
  • Clone number
  • Isotype
  • Research areas


Our Abpromise guarantee covers the use of ab38689 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr 1/500.
ELISA 1/1000.
WB 1/4000. Predicted molecular weight: 27 kDa.
IHC-P 1/50 - 1/100.


  • Relevance
    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 Ca2+ -activated photoprotein aequorin.

    Subunit structure: Monomer.

    Tissue specificity: Photocytes.

    Post-translational modification: Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.

    Biotechnological use: Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.

    Sequence similarities: Belongs to the GFP family.

    Biophysicochemical properties: Absorption: Abs(max)=395 nm
    Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm.
  • Alternative names
    • GFP antibody
    • Green fluorescent protein antibody


  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse skin tissue labelling GFP with ab38689. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 38°C; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A peroxidase-conjugated goat anti-mouse polyclonal (ready to use) was used as the secondary antibody. -ve shows negative staining of the antibody in a wild type mouse, whereas +ve shows positive staining in a mouse expressing GFP under the LGR5 promoter.

  • Immunohistochemistry (Frozen sections) analysis of E10.5 mouse embryo tissue sections labeling GFP with ab38689 at 1/200 dilution. The tissue was fixed with paraformaldehyde and permeabilized with PBS / 0.5% v/v Triton X-100. The tissue was incubated with ab38689 in 1% BSA + 5% goat serum + PBST for 12 hours at 25°C. A polyclonal goat anti-mouse Alexa Fluor® 568 secondary antibody was used at 1/1000 dilution.

    See Abreview

  • Anti-GFP antibody [6AT316] (ab38689) at 1/4000 dilution + GFP recombinant protein lysate at 35 µg

    Peroxidase-conjugated goat anti-mouse IgG (H+L) at 1/5000 dilution

    Predicted band size: 27 kDa

    Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-GFP antibody [6AT316] (ab38689)

    Lane 1 : GFP at 1 µg
    Lane 2 : GFP at 0.1 µg
    Lane 3 : GFP at 0.01 µg
    Lane 4 : YFP at 1 µg
    Lane 5 : YFP at 0.1 µg
    Lane 6 : YFP at 0.01 µg

    Predicted band size: 27 kDa
    Observed band size: 27 kDa

    GFP and YFP were expressed in bacteria.
  • Formalin-fixed and paraffin-embedded human cancer tissue reacted with ab38689, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining.
  • ab38689 staining GFP in Chicken spinal cord tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500 in PBS) for 16 hours at 4°C. A Cy2®-conjugated Donkey anti-mouse monoclonal (1/100) was used as the secondary antibody.

    See Abreview


This product has been referenced in:
  • Mitra S  et al. Dual regulation of lin28a by Myc is necessary during zebrafish retina regeneration. J Cell Biol 218:489-507 (2019). Read more (PubMed: 30606747) »
  • Schiller LT  et al. Enhanced Production of Exosome-Associated AAV by Overexpression of the Tetraspanin CD9. Mol Ther Methods Clin Dev 9:278-287 (2018). Read more (PubMed: 29707602) »
See all 20 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Mouse Tissue sections (pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C

Abcam user community

Verified customer

Submitted Jul 13 2017

Human Cell lysate - whole cell (HeLa Cells)
Total protein in input
1e+007 cells
Transfected with EGFP 48 hrs
Immuno-precipitation step
Protein G
HeLa Cells

Abcam user community

Verified customer

Submitted Oct 20 2015

Immunohistochemistry (Frozen sections)
Blocking step
NOVOCASTRA protein block as blocking agent as blocking agent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Mouse Tissue sections (E10.5 mouse embryo)
E10.5 mouse embryo
Yes - PBS / 0.5% v/v Triton X-100 (Sigma)

Abcam user community

Verified customer

Submitted Mar 20 2014


The epitope of the antibody has not been mapped, and it has not been tested with eGFP, so we cannot guarantee it detects eGFP. However, EGFP differs from GFP by only a few amino acids, there is a great chance the antibody will detect eGFP.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Immunohistochemistry (Frozen sections)
Chicken Tissue sections (Spinal cord)
Spinal cord
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Dr. A Klar

Verified customer

Submitted Apr 12 2011


PROCEDURE: Solution Preparation PBS, pH7.4: 10 mM Na2HPO4, 1.8 mM KH2PO4, 50 mM NaCl, 2.7 mM KCl Blocking buffers: 5% BSA/0.5% Tween-20 in 1X PBS. Protocol 1. Deparaffinize and rehydrate sections as follows: 3 x 3 min with xylene; 3 x 2 min with 100% ethanol; 2 min with 95% ethanol, 2 min with 80% ethanol, 2 min with 70% ethanol, and 5 min with PBS. 2. Perform citrate buffer antigen retrieval (see below). 3. Block endogenous peroxidases by soaking slides in a solution of 90% methanol/3% H2O2 for 15 min at room temperature (RT). Then wash 3 x 5 min with PBS. 4. Shake and wipe off excess PBS. Circle all sections with a pap pen. Add 75 ul of Blocking buffer to each section immediately. Do not touch sections with tip. 5. Incubate 1 hr to overnight at RT in a humidified chamber. Do not let the slides touch each others. 6. Dilute primary antibody in Blocking buffer. Add 75 ul per section and incubate 1 hr to overnight at RT in a humidified chamber. 7. Drain primary antibody off section. Wash slides 3 x 10 min in PBS. You may have to wash slides in PBS + 0.1%-0.5% Tween-20 for some primary antibodies. 8. Dilute secondary antibody 1:1000 (this may require optimisation) in Blocking buffer without Tween-20. Add 75 ul per section and incubate for 1 hr at RT in a humidified chamber. 9. Drain secondary antibody and wash slides 5 x 10 min in PBS + 0.1% Tween-20. (For secondary antibodies that are peroxidase conjugated, go to step 11.) 10. Make ABC according to manufacturers instructions 30 min before time of use (mix 5 ml of PBS with 2 drops of solution A and 2 drops of solution B). Incubate samples for 45 min at RT. Wash 5 min in PBS. 11. Make DAB solution according to manufacturers instructions. WEAR GLOVES: Mix 5 ml ddH2O with 2 drops of buffer, 4 drops of DAB and 2 drops of H2O2. (If you want a gray-black stain, add 2 drops of the Nickel solution, and mix). Add immediately to slides and wait for color change (approximately 2-10 min). Drain slides and place into ddH2O for 5 min. Dispose of DAB waste with bleach. 12. Counterstain with methyl green (1 min) or hematoxylin (3 sec). Wash 3 times with ddH2O. 13. Immediately dehydrate in 70% ethanol, 80% ethanol, and 100% ethanol (one dip each). 14. Mount and seal coverslip with nail polish. This ab38689 GFP antibody ab38689 works well with Sodium Citrate Antigen Retrieval: Sodium Citrate Antigen Retrieval: 1. Place slides in a glass slide holder and fill in the rest of the rack with blank slides (10 total) to ensure even heating. 2. Place rack in 600 ml of 10 mM sodium citrate, pH 6.0 in a glass 2L-beaker. Mark a line at the top of the liquid on the beaker. 3. Microwave for 20 min total, replacing evaporated water every 5 min. 4. Cool slides for 20 min. 5. Wash 4 x 3 min in ddH2O, and 3 min in 1X PBS.

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