GFP Antibody

Catalog # Availability Size / Price Qty
AF4240
AF4240-SP
Detection of GFP by Western Blot.
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Product Details
Citations (17)
FAQs
Supplemental Products
Reviews (3)

GFP Antibody Summary

Specificity
Detects GFP in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant GFPuv
Ser2-Lys238
Accession # P42212
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
2.5 µg/mL
See below
Immunoprecipitation
25 µg/mL
Conditioned cell culture medium spiked with Recombinant GFP, see our available Western blot detection antibodies
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
1-15 µg/mL
See below
Intracellular Staining by Flow Cytometry
0.25 µg/106 cells
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of GFP antibody by Western Blot. View Larger

Detection of GFP by Western Blot. Western blot shows lysates of NS0 mouse myeloma cell line either mock transfected or transfected with eGFP-tagged EDG6. PVDF membrane was probed with 1 µg/mL of Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for GFP at approximately 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry GFP antibody in HEK293 human embryonic kidney cells transfected with GFP by Immunocytochemistry (ICC). View Larger

GFP in HEK293 human embryonic kidney cells transfected with GFP. GFP was detected in immersion fixed HEK293 human embryonic kidney cells transfected with GFP (green) using Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, middle panel; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm of GFP-positive cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Intracellular Staining by Flow Cytometry Detection of GFP antibody in HEK293 human embryonic kidney cells transfected with GFP. View Larger

Detection of GFP in HEK293 human embryonic kidney cells transfected with GFP. HEK293 human embryonic kidney cells transfected with GFP was stained with Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody only (A, Catalog # F0108) or with Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) followed by Secondary Antibody (B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. Quadrant markers were set based on control antibody staining (Catalog # AB-108-C).

Simple Western Detection of GFP antibody by Simple Western<sup>TM</sup>. View Larger

Detection of GFP by Simple WesternTM. Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line either mock transfected (-) or transfected with eGFP-tagged EDG6 (+), loaded at 0.2 mg/mL. A specific band was detected for GFP at approximately 114 kDa (as indicated) using 2.5 µg/mL of Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Simple Western GFP Antibody View Larger

Simple Western: GFP Antibody [Unconjugated] [AF4240] - Simple Western: GFP Antibody [Unconjugated] [AF4240] - Design for a membrane-localized ACE2 expression system. (A) Our ACE2 construct is driven by a CMV promoter followed by the first 25 residues of ACE2 containing the leader sequence that direct ACE2 to the plasma membrane. This is followed by a 3xHA tag linked to the remainder of ACE2 (20-805) and a C-terminal sfGFP. Both 3xHA and sfGFP fusions are separated from ACE2 by flexible 3xGGGGS linkers. (B) The ACE2 fusion protein is designed to be embedded in the plasma membrane where it can perform extracellular carboxypeptidase-mediated metabolism and its levels can be detected by cell staining with antibodies to HA. (C) Lysates from untransfected or 3xHA-ACE2-sfGFP-transfected HEK293 cells were analyzed by automated Jess capillary immunoassay using antibodies to HA, GFP, and two ACE2 antibodies. (D) Confocal fluorescence microscopy of HEK cells transfected with 3xHA-ACE2-sfGFP and stained with HA and the nuclear stain DAPI. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/37644110), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: GFP

Green fluorescent protein (GFP) is a 27 kDa protein originally isolated from the jellyfish Aequorea victoria. In the presence of UV light (490-520 nm), it emits a green fluorescent color that can be used to pinpoint locations of various intracellular proteins. GFP is 238 amino acids (aa) in length. It is a globular monomer that has a tendency to dimerize. The monomer has the shape of a beta -barrel with a chromophore (aa 65-67) containing alpha -helix running up its center. GFPuv is the Aequorea sequence with three aa substitutions; Phe to Ser at # 99, Met to Thr at # 153, and Val to Ala at # 163. This form expresses faster and is 18-fold brighter than native GFP; excitation peaks at 395 nm and emission at 508 nm.

Long Name
Green Fluorescent Protein
Alternate Names
eGFP; Enhanced Green Fluorescent Protein; GFP; GFPuv; green fluorescent protein (gfp); Green Fluorescent Protein

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Citations for GFP Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

17 Citations: Showing 1 - 10
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  1. Smooth Muscle Cell Reprogramming in Aortic Aneurysms
    Authors: Chen PY, Qin L, Li G et al.
    Cell Stem Cell
  2. A cell-based assay for rapid assessment of ACE2 catalytic function
    Authors: Warren M. Meyers, Ryan J. Hong, Wun Chey Sin, Christine S. Kim, Kurt Haas
    Scientific Reports
  3. Proteomics uncover EPHA2 as a potential novel therapeutic target in colorectal cancer cell lines with acquired cetuximab resistance
    Authors: Lucien Torlot, Anna Jarzab, Johanna Albert, Ágnes Pók-Udvari, Arndt Stahler, Julian Walter Holch et al.
    Journal of Cancer Research and Clinical Oncology
  4. Blood-brain barrier penetration of non-replicating SARS-CoV-2 and S1 Variants of Concern induce neuroinflammation which is accentuated in a mouse model of Alzheimer's disease
    Authors: MA Erickson, AF Logsdon, EM Rhea, KM Hansen, SJ Holden, WA Banks, JL Smith, C German, SA Farr, JE Morley, RR Weaver, AJ Hirsch, A Kovac, E Kontsekova, KK Baumann, MA Omer, J Raber
    Brain, Behavior, and Immunity, 2023-01-20;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  5. Extending the dynamic range of biomarker quantification through molecular equalization
    Authors: Newman, S;Wilson, B;Mamerow, D;Wollant, B;Nyein, H;Rosenberg-Hasson, Y;Maecker, H;Eisenstein, M;Soh, H;
    bioRxiv
    Species: Human
    Sample Types: Serum
    Applications: Bioassay
  6. Intestinal inflammation and increased intestinal permeability in Plasmodium chabaudi AS infected mice
    Authors: Jason P Mooney, Sophia M DonVito, Rivka Lim, Marianne Keith, Lia Pickles, Eleanor A Maguire et al.
    Wellcome Open Research
  7. ESCRT-I fuels lysosomal degradation to restrict TFEB/TFE3 signaling via the Rag-mTORC1 pathway
    Authors: Marta Wróbel, Jarosław Cendrowski, Ewelina Szymańska, Malwina Grębowicz-Maciukiewicz, Noga Budick-Harmelin, Matylda Macias et al.
    Life Science Alliance
  8. MSC Pretreatment for Improved Transplantation Viability Results in Improved Ventricular Function in Infarcted Hearts
    Authors: MF Pittenger, S Eghtesad, PG Sanchez, X Liu, Z Wu, L Chen, BP Griffith
    International Journal of Molecular Sciences, 2022-01-08;23(2):.
    Species: Rat
    Sample Types: Cell Lysates
    Applications: Western Blot
  9. Evidence of a Myenteric Plexus Barrier and Its Macrophage-Dependent Degradation During Murine Colitis: Implications in Enteric Neuroinflammation
    Authors: D Dora, S Ferenczi, R Stavely, VE Toth, ZV Varga, T Kovacs, I Bodi, R Hotta, KJ Kovacs, AM Goldstein, N Nagy
    Cellular and Molecular Gastroenterology and Hepatology, 2021-07-08;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  10. Neonatal diabetes mutations disrupt a chromatin pioneering function that activates the human insulin gene
    Authors: I Akerman, MA Maestro, E De Franco, V Grau, S Flanagan, J García-Hur, G Mittler, P Ravassard, L Piemonti, S Ellard, AT Hattersley, J Ferrer
    Cell Reports, 2021-04-13;35(2):108981.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  11. Wnt/Beta-catenin/Esrrb signalling controls the tissue-scale reorganization and maintenance of the pluripotent lineage during murine embryonic diapause
    Authors: R Fan, YS Kim, J Wu, R Chen, D Zeuschner, K Mildner, K Adachi, G Wu, S Galatidou, J Li, HR Schöler, SA Leidel, I Bedzhov
    Nat Commun, 2020-10-30;11(1):5499.
    Species: Mouse
    Sample Types: Whole Cells, Whole Tissue
    Applications: ICC, IHC
  12. Molecular Motor KIF3B Acts as a Key Regulator of Dendritic Architecture in Cortical Neurons
    Authors: Nadine F. Joseph, Eddie Grinman, Supriya Swarnkar, Sathyanarayanan V. Puthanveettil
    Frontiers in Cellular Neuroscience
  13. Splicing variation of BMP2K balances abundance of COPII assemblies and autophagic degradation in erythroid cells
    Authors: J Cendrowski, M Kaczmarek, M Mazur, K Kuzmicz-Ko, K Jastrzebsk, M Brewinska-, A Kominek, K Piwocka, M Miaczynska
    Elife, 2020-08-14;9(0):.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: Co-IP, ICC, Western Blot
  14. Agrin has a pathological role in the progression of oral cancer
    Authors: C Rivera, FS Zandonadi, C Sánchez-Ro, CD Soares, DC Granato, WA González-A, AF Paes Leme
    Br. J. Cancer, 2018-06-06;0(0):.
    Applications: Immunoprecipitation
  15. Natural killer cells attenuate cytomegalovirus-induced hearing loss in mice
    Authors: Ali A. Almishaal, Pranav D. Mathur, Elaine Hillas, Liting Chen, Anne Zhang, Jun Yang et al.
    PLOS Pathogens
  16. Lymphatic endothelial progenitors originate from plastic myeloid cells activated by toll-like receptor-4
    Authors: LD Volk-Drape, KL Hall, AC Wilber, S Ran
    PLoS ONE, 2017-06-09;12(6):e0179257.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  17. Efficient Generation of Cardiac Purkinje Cells from ESCs by Activating cAMP Signaling
    Authors: Su-Yi Tsai, Karen Maass, Jia Lu, Glenn I. Fishman, Shuibing Chen, Todd Evans
    Stem Cell Reports

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Reviews for GFP Antibody

Average Rating: 4 (Based on 3 Reviews)

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GFP Antibody
By Anonymous on 09/17/2018
Application: Immunocytochemistry/Immunofluorescence Sample Tested: Brain (cross-section through blood vessel) Species: Mouse

GFP Antibody
By Anonymous on 01/17/2018
Application: IP Sample Tested: HEK293 human embryonic kidney cell line Species: Human

GFP Antibody
By Anonymous on 10/27/2017
Application: WB Sample Tested: Liver cells Species: Mouse