Name: Anti-GFP antibody [EPR14104]
SKU: ab183734
Rating: 5 (24 reviews)

Anti-GFP antibody [EPR14104] (ab183734) is a rabbit monoclonal antibody detecting GFP in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF.

- Biophysical QC for unrivalled batch-batch consistency

- Over 80 publications

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (AB183734), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	IHC-P	WB	ICC/IF	Flow Cyt (Intra)
Tag	Not recommended	Tested	Tested	Tested	Tested

Not recommended
Not recommended

Species	Dilution info	Notes
Species Tag	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Tag	Dilution info 1/100 - 1/250	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Tag	Dilution info 1/10000 - 1/50000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Tag	Dilution info 1/500	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Tag	Dilution info 1/50	Notes -

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Target data

See full target information GFP

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

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Clone number

EPR14104

Purification technique

Affinity purification Protein A

Specificity

On the basis of low sequence homology, ab183734 is predicted to show no or limited cross-reactivity to RFP and BFP.

For the best results in IHC, please use ab183734 on Formalin/PFA-fixed paraffin-embedded sections with heat mediated antigen retrieval.

Dissociation constant

1.11 x 10^-11 M

Concentration

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

Anti-GFP antibody [EPR14104] (ab183734) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P and WB.

Abcam's high quality manufacturing and validation processes ensure Anti-GFP antibody [EPR14104] (ab183734) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

Anti-GFP antibody [EPR14104] (ab183734) has 24 independent reviews from customers.

GFP antibodies are often used as loading controls in Western Blot. Anti-GFP antibody [EPR14104] has been verified in Western Blot samples and detects a band at 27kDa Molecular weight.

GFP antibodies are used to visualize proteins labelled with this tag in a variety of applications (for example imaging and Flow cytometry). To enable specific detection of your tagged protein, Anti-GFP antibody [EPR14104] (ab183734) has been validated in Flow Cyt (Intra), ICC/IF, IHC-P and WB.

Anti-GFP antibody [EPR14104] (ab183734) specifically detects GFP (UniProt ID: P42212; Molecular weight: 27kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPR14104 - Anti-GFP antibody [EPR14104] - BSA and Azide free ab220802.

Antibody clone EPR14104 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 488, APC, PE (Alexa Fluor® 488 Anti-GFP antibody [EPR14104] - GFP booster ab225314, APC Anti-GFP antibody [EPR14104] ab283597, PE Anti-GFP antibody [EPR14104] ab303588).

Green Fluorescent Protein (GFP), originally derived from the jellyfish Aequorea victoria, emits a bright green fluorescence under ultraviolet or blue light. This unique property makes GFP an invaluable tool in molecular and cell biology research. Widely used to tag proteins, GFP allows scientists to visualize and track protein expression, localization and interactions within living cells. Key applications of GFP include fluorescence microscopy (ICC, IHC, IF), flow cytometry (FC) and western blotting (WB).

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

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.

12 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (ab183734)
Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse liver tissue (left) and normal mouse liver tissue (right) labeling GFP with unpurified ab183734 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (ab183734)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of GFP transgenic mouse pancreas tissue labelling GFP with purified ab183734 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] (ab183734)
ab183734 staining GFP in GFP-transfected NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab183734 at 1/500 dilution overnight at +4°C followed by incubation with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488), for 1 hour, at 1μg/ml.
Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH3T3 cells, the cells upon which ab183734 was applied gave a stronger signal in the 488 channel, indicating that ab183734 is binding to GFP and therefore eliciting signal amplification.
ab183734 was also applied to non-GFP-transfected NIH3T3 cells, which produced no positive staining, indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue).
Western blot - Anti-GFP antibody [EPR14104] (ab183734)
Blocking and dilution buffer: 5% NFDM /TBST.
All lanes: Western blot - Anti-GFP antibody [EPR14104] (ab183734) at 1/10000 dilution
Lane 1: GFP transfected 293 cell lysate at 10 µg
Lane 2: Non-transfected 293 cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 27 kDa
Observed band size: 25 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (ab183734)
Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse colon tissue (left) and normal mouse colon tissue (right) labeling GFP with unpurified ab183734 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] (ab183734)
Immunofluorescent analysis of 4% paraformaldehyde-fixed GFP transfected 293 cells labeling GFP with unpurified ab183734 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody at 1/200 dilution (green). Counterstained with Dapi (blue)
Western blot - Anti-GFP antibody [EPR14104] (ab183734)
Different batches of ab183734 were tested on GFP transfected HEK-293 (Human embryonic kidney epithelial cell) lysate at 0.004 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 27 kDa.
All lanes: Western blot - Anti-GFP antibody [EPR14104] (ab183734)
Predicted band size: 27 kDa
Western blot - Anti-GFP antibody [EPR14104] (ab183734)
All lanes: Western blot - Anti-GFP antibody [EPR14104] (ab183734) at 1/10000 dilution
Lane 1: GFP transfected 293 cell lysate at 10 µg
Lane 2: Non-transfected 293 cell lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (ab183734)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse pancreas tissue (negative control) labelling GFP with purified ab183734 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-GFP antibody [EPR14104] (ab183734)
All lanes: Western blot - Anti-GFP antibody [EPR28018-38] (Anti-GFP antibody [EPR28018-38] ab314656) at 1/1000 dilution
Lane 1: 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 7 µg
Lane 2: 293T transfected with an empty vector containing an EGFP-myc-tag, whole cell lysate at 7 µg
Lane 3: 293T transfected with an empty vector containing a GFP-tag, whole cell lysate at 7 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 26 kDa, 36 kDa
Exposure time: 3s
Flow Cytometry (Intracellular) - Anti-GFP antibody [EPR14104] (ab183734)
Intracellular Flow Cytometry analysis of 293T (human embryonic kidney) transfected with human TNFRSF9 cells labeling GFP with purified ab183734 at 1/50 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 647) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
This image is courtesy of a customer review submitted by Estela Munoz
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (ab183734)
Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed, mouse pineal gland tissue from an adult Cx3cr1-eGFP+/-; APP/PS1 mouse. Stained with ab183734 at 1/300 dilution. Secondary antibody used was Alexa Fluor® 647 donkey Anti-rabbit antibody at 1/1000 dilution. Cell nuclei were counterstained with DAPI (grey). Blocking was done with 3% BSA for 1 hour at 24°C. The sample was incubated with the primary antibody and PBS with 3% BSA and 1% Triton X-100 for 16 hours at 24°C. Antigen retrieval method was heat mediated 10 mM citrate buffer pH6