GFP Antibody - BSA Free

Novus Biologicals | Catalog # NB600-303

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Non-species specific

Cited:

Jellyfish

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Electron Microscopy

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Chemotaxis, IF/IHC, Electron Microscopy

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all GFP Primary Antibodies »

Product Specifications

Immunogen

The immunogen is a Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence (246aa) derived from the jellyfish Aequorea victoria.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for GFP Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-303]

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-303]

Immunocytochemistry/Immunofluorescence: GFP Antibody [NB600-303] - Analysis of FITC conjugate of NB600-303. Staining of mouse spleen cells Tissue: spleen cells infected with MHV68-H2bYFP. As seen in: Collins CM, Speck SH (2012) Tracking Murine Gammaherpesvirus 68 Infection of Germinal Center B Cells.
Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303] - Analysis using the FITC conjugate of NB600-303. Detection of Lane 1: GFP. Load: 50 ug per lane. Primary antibody: none. Secondary antibody: Fluorescein conjugated Anti-GFP at 1:1000 for 60 min at RT. Block: 1% BSA-TTBS for 30 min at RT. Predicted/Observed size: 28 kDa, 28 kDa.
Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303] - WB analysis of GFP in various lysates. Image courtesy of product review by Emilie Leclerc.
Immunoprecipitation: GFP Antibody [NB600-303]

Immunoprecipitation: GFP Antibody [NB600-303]

Immunoprecipitation: GFP Antibody [NB600-303] - IP analysis of GFP in HeLa cell lysate. Image courtesy of anonymous customer review.
Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303] - Analysis of GFP HRP expression in HeLa/GFP cell lysate.
Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303] - Analysis using Biotin conjugate of NB600-303. Lane 1: 50ng of GFP. Lane 2: none. Primary antibody: none. Secondary antibody: Anti- Biotin Conjugated secondary antibody was used at 1:5000 in Blocking Buffer for Fluorescent Western Blotting for 45 min at RT. HRP Streptavidin was used at 1:40,000 in incubated with blocking buffer for 30 min at 20C. Block: 5% Blotto 30 min at 20C. Predicted/Observed size: 28 kDa for GFP. Other band(s): none.
Immunohistochemistry: GFP Antibody [NB600-303]

Immunohistochemistry: GFP Antibody [NB600-303]

GFP-Antibody-Immunohistochemistry-NB600-303-img0024.jpg
Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303]

Western Blot: GFP Antibody [NB600-303] - Lane 1: 293FT cells transfected with CDK4 dominant negative. Lane 2: 293FT cells poitive control. Load: 25 ug per lane. Primary antibody: GFP antibody at 1:400 for overnight at 4C. Secondary antibody: IRDye800 (TM) rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 27 kDa for GFP.
Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-303]

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-303]

Immunocytochemistry/Immunofluorescence: GFP Antibody [NB600-303]
GFP Antibody

GFP Antibody

Western Blot of GFP Antibody. Lane 1: 293FT cells transfected with CDK4 dominant negative. Lane 2: 293FT cells poitive control. Load: 25 ug per lane. Primary antibody: GFP antibody at 1:400 for overnight at 4C. Secondary antibody: IRDye800(TM) rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 27 kDa for GFP.
GFP Antibody

GFP Antibody

Western Blot of Rabbit anti-GFP antibody. Lane 1: Wild type GFP (0.1 ug) was used to spike HeLa whole cell lysate. Lane 2: none. Load: 30 ug per lane. Primary antibody: GFP antibody at 1:1000 for overnight at 4C. Secondary antibody: IRDye800(TM) Goat-a-Rabbit IgG [H&L] MX10 () at 1:10,000 for 45 min at RT. Block: 5% BLOTTO in PBS overnight at 4C. Predicted/Observed size: 27 kDa for epitope tag GFP. Other band(s): none.
GFP Antibody

Western Blot: GFP Antibody [NB600-303] -

The polo-boxes of Plk4 are not sufficient to mediate STIL binding.(A) Scheme of Flag-Plk4 fragments. (B) After overexpression of Flag-Plk4-fragments (A) and GFP-STIL in HEK293T cells, cell lysates were subjected to immunoprecipitations using anti-Flag antibodies. Coprecipitation of GFP-STIL with Flag-Plk4 fragments was detected by western blotting using anti-GFP and anti-Flag antibodies.
GFP Antibody

Western Blot: GFP Antibody [NB600-303] -

Western Blot: GFP Antibody [NB600-303] - The polo-boxes of Plk4 are not sufficient to mediate STIL binding.(A) Scheme of Flag-Plk4 fragments. (B) After overexpression of Flag-Plk4-fragments (A) & GFP-STIL in HEK293T cells, cell lysates were subjected to immunoprecipitations using anti-Flag antibodies. Coprecipitation of GFP-STIL with Flag-Plk4 fragments was detected by western blotting using anti-GFP & anti-Flag antibodies. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/4/3/370/1390/Plk4-dependent…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
GFP Antibody

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-303] -

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-303] - Lgr5+ liver stem cells transplantation decreased liver fibrosis. a Schematic overview of the experimental setup. Eight-week-old wild-type C57 mice were i.p. injected with CCL4 (2 ml/kg, Sigma-Aldrich) dissolved in olive oil at a ratio of 1:4, or olive oil alone (2 ml/kg) twice a week for 6 weeks. Lgr5-GFP+ liver stem cells or primary hepatocyte (PH) derived from Lgr5-GFP mice were transplanted into the liver fibrosis mice by intrasplenical injection on day 0. b Lgr5+ cells were isolated from Lgr5-GFP mice treated with CCL4 by FACS assay for transplantation (left). The liver Lgr5 expression was stained using anti-GFP antibody in mice with Lgr5+ cells or PH transplantation (right) on day 40. n = 10 mice, scale bars, 200 μm. c, d Lgr5+ liver stem cells transplantation decreased CCL4-induced liver fibrosis & recovered liver functions. Lgr5-GFP+ liver stem cells or PH transplantation were described in a; the livers were harvested & stained using H&E & Sirius Red for fibrosis analysis & quantification of positive-staining areas measured by Image J software (c). The serum was harvested for ALT & AST analysis (d, e). For c, scale bars, 200 μm, the results are shown as mean ± s.d. of five independent sections taken randomly per mice & a total of 50 tissue specimens in each group (n = 10 mice) *p < 0.05, **p < 0.01. For d, e, triplicates for each condition were analyzed. The results are shown as mean ± s.d. of three independent experiments. *p < 0.05, **p < 0.01 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29079780), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for GFP Antibody - BSA Free

Application
Recommended Usage

ELISA

1:10000-1:40000

Flow Cytometry

1:10 - 1:1000

Immunocytochemistry/ Immunofluorescence

1:10 - 1:500

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Frozen

1:10 - 1:500

Immunohistochemistry-Paraffin

1:10 - 1:500

Immunoprecipitation

1:10 - 1:500

Simple Western

1:500

Western Blot

1:1000-1:3000
Application Notes

This product has been tested in ELISA and western blot. Expect ~27kDa band. Although not tested, this antibody is suitable for IP, IF, ChIP, and Flow. Specific conditions for reactivity should be optimized by the end user.

Immunoprecipitation reported successful by customer review.

Immunohistochemistry-Paraffin (PMID: 21311774) Immunocytochemistry/Immunofluorescence was reported in scientific literature.

Use in Flow Cytometry reported in scientific literature (PMID: 23903657).

Use in Electron Microscopy reported in scientific literature (PMID 25538186).

Use in Immunohistochemistry-Frozen reported in multiple pieces of scientific literature.

Use in chromatin immunoprecipitation reported in scientific literature (PMID 28174757).
See Simple Western Antibody Database for Simple Western validation: tested in human melanoma cells; antibody dilution of 1:500; separated by size

Reviewed Applications

Read 3 reviews rated 4 using NB600-303 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Format

BSA Free

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: GFP

Green fluorescent protein (GFP) is a 27 kDa protein derived from the bioluminescent jellyfish Aquorea Victoria. GFP can be excited by light in the blue to ultraviolet spectrum and has an excitation peak at 395 nm with an emission peak at 509 nm in the green spectrum. GFP autocatalytically forms a fluorescent pigment from natural amino acids present in the nascent protein. GFP is widely used as a reporter (tag) for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. GFP fluorescence is stable under fixation conditions and antibodies are suitable for a variety of applications including ELISA, ICC/IF, IHC, IP and WB (1-3).

References
1. Shi, C., Pan, F. C., Kim, J. N., Washington, M. K., Padmanabhan, C., Meyer, C. T.,... Means, A. L. (2019). Differential Cell Susceptibilities to Kras(G12D) in the Setting of Obstructive Chronic Pancreatitis. Cell Mol Gastroenterol Hepatol. doi:10.1016/j.jcmgh.2019.07.001

2. Zhao, S., Fortier, T. M., & Baehrecke, E. H. (2018). Autophagy Promotes Tumor-like Stem Cell Niche Occupancy. Curr Biol, 28(19), 3056-3064.e3053. doi:10.1016/j.cub.2018.07.075

3. Zusso, M., Lunardi, V., Franceschini, D., Pagetta, A., Lo, R., Stifani, S.,... Moro, S. (2019). Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway. J Neuroinflammation, 16(1), 148. doi:10.1186/s12974-019-1538-9

Long Name

Green Fluorescent Protein

Alternate Names

eGFP, GFPuv

Additional GFP Products

Product Documents for GFP Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for GFP Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for GFP Antibody - BSA Free

Customer Reviews for GFP Antibody - BSA Free (3)

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Hela whole cell lysate
    Species: Human
    Verified Customer | Posted 11/18/2011
    GFP Antibody - BSA Free NB600-303
  • Name: Marisa Carbonaro
    Application: Immunoprecipitation
    Sample Tested: HeLa cells
    Species: Human
    Verified Customer | Posted 09/27/2010
    GFP Antibody - BSA Free NB600-303
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: recombinant protein, Sample Amount: 0.5 - 1 ug
    Species: Other
    Verified Customer | Posted 03/02/2009

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for GFP Antibody - BSA Free

Showing  1 - 4 of 4 FAQs Showing All

    • A: The current lot (31271) of NB600-303 is provided at a concentration of 6.05mg/ml.

    • A: NB600-303 has been validated for detection of S65T-GFP, RS-GFP, YFP and EGFP. Based on an alignment to a ECFP sequence that I found on NCBI (ncbi site), the GFP protein this product was raised against does show a good amount of homology. Therefore, there would be a good chance for cross reaction of this product against ECFP.

    • A: Both antibodies (i.e. # ab290 from Abcam and # NB600-303 from Novus) use the same immunogen to develop this antibody. Both companies used highly purified recombinant full length protein made in E. coli as immunogen and this antibody is directed against the entire GFP molecule. We stand behind our products 100% and would be there to help in case you comes up with any concerns. Moreover our antibody is also well tested and has been cited in recent research papers in journals of high repute (Dis Model Mech. 2011 May;4(3):359-67; PLoS ONE. 2011; J Cell Sci. 2010 May 15;123(Pt 10):1732-41; Acta Anat (Basel). 1991;141(1):82-4; FASEB J. 2009 Nov 25. and many more...).

    • Q: We are looking for a rabbit anti-GFP antibody as a primary antibody for immunofluorescence and WB. I've seen that you carry several anti-GFP antibodies and I would like to ask which one you would recommend. The one with Cat.No NB600-308 has been used in a lot of publications and I reckon it is a solid antibody. Is this our best choice, or is there a better alternative?
      A: NB600-308 is an excellent GFP antibody and I think it would work very well for you. It has been reviewed and published with which usually makes our customer feel much more confident, seeing as it worked in other people's hand.
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