GFP Antibody (9F9.F9) - BSA Free

Novus Biologicals | Catalog # NB600-597

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Non-species specific

Cited:

Human

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunoblotting, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Dot Blot, In vitro assay, Knockdown Validated

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunoblotting, ELISA, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Cytometric Bead Assay Standard, In vitro assay, IF/IHC, Knockdown Validated

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 9F9.F9

Format

BSA Free
View all GFP Primary Antibodies »

Product Specifications

Immunogen

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

Reactivity Notes

Reactivity is observed against recombinant Green Fluorescent Protein from Aequorea victoria by both Western blot and ELISA. No reaction is seen against RFP.
Recognizes fusion proteins in Drosophila, reactivity reported in scientific literature (PMID: 26586808). Human reactivity reported in scientific literature (PMID: 28198010).

Specificity

Reactivity is observed against recombinant Green Fluorescent Protein from Aequorea victoria by both Western blot and ELISA. No reaction is seen against RFP.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Description

GFP Monoclonal Antibody was prepared from tissue culture supernatant by Protein A affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against anti-Mouse Serum

Store this antibody 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 (9F9.F9) - BSA Free

Immunocytochemistry/ Immunofluorescence: GFP Antibody (9F9.F9) [NB600-597]

Immunocytochemistry/ Immunofluorescence: GFP Antibody (9F9.F9) [NB600-597]

Immunocytochemistry/Immunofluorescence: GFP Antibody (9F9.F9) [NB600-597] - GFP-transducted 293T cells. Image from verified customer review.
Western Blot: GFP Antibody (9F9.F9) [NB600-597]

Western Blot: GFP Antibody (9F9.F9) [NB600-597]

Western Blot: GFP Antibody (9F9.F9) [NB600-597] - Lane 1: Molecular Weight Markers. Lane 2: GFP. Load: 10ng per lane. Primary antibody: Ms Anti-GFP antibody at 1:1000 for overnight at 4C. Secondary antibody: Mouse HRP secondary antibody at 1:40,000 for 30 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 27 kDa, 27 kDa for epitope tag GFP. Other band(s): None.
Western Blot: GFP Antibody (9F9.F9) [NB600-597]

Western Blot: GFP Antibody (9F9.F9) [NB600-597]

Western Blot: GFP Antibody (9F9.F9) [NB600-597] - All lanes have picogram amount recombinant GFP protein spiked into HeLa cell-derived lysates Lane 1: 64pg 2pg Lane 3: 16pg Lane 4: 8pg Lane 5: 4pg Lane 6: 2pg Lane 7: 1g Lane 8: 0pg. Primary antibody: anti-GFP monoclonal antibody at 1:400 for overnight at 4C. Secondary antibody: HRP-conjugated anti-Mouse IgG was performed at a dilution of 1:20,000 for 1h at 4C. Block: TTBS supplemented with 1% BSA for 1 h at 4C. Predicted/Observed size: 27 kDa for GFP. Other b&(s): none.
Immunohistochemistry: GFP Antibody (9F9.F9) [NB600-597]

Immunohistochemistry: GFP Antibody (9F9.F9) [NB600-597]

Immunohistochemistry: GFP Antibody (9F9.F9) [NB600-597] - Used 1:5000 As referenced in: Lu S-M, Tremblay M-E`, King IL, Qi J, Reynolds HM, et al. (2011) HIV-1 Tat-Induced Microgliosis and Synaptic Damage via Interactions between Peripheral and Central Myeloid Cells. PLoS ONE 6(9): e23915. doi:10.1371/journal.pone.0023915 Image using the Biotin format of this antibody.
Western Blot: GFP Antibody (9F9.F9) [NB600-597]

Western Blot: GFP Antibody (9F9.F9) [NB600-597]

Western Blot: GFP Antibody (9F9.F9) [NB600-597] - Expression of GFP in CHO-K1 cells trasfected with GFP. Image from verified customer review.
GFP Antibody (9F9.F9)

GFP Antibody (9F9.F9)

Immunoprecipitation/Western Blot using GFP Protein. Lane 1: Opal Prestained Molecular Weight Marker
GFP Antibody (9F9.F9)

GFP Antibody (9F9.F9)

Western blot of GFP Antibody (9F9.F9). Lane 1: Opal Prestained Molecular Weight Marker
GFP Antibody (9F9.F9)

GFP Antibody (9F9.F9)

Western blot of Mouse Anti-GFP Antibody. Lane 1: Thermo SuperSignal Molecular Weight Marker. Lane 2: GFP protein

Applications for GFP Antibody (9F9.F9) - BSA Free

Application
Recommended Usage

Dot Blot

1:100 - 1:2000

ELISA

1:10000-1:30000

Flow Cytometry

1:1:10 - 1:1000

Immunocytochemistry/ Immunofluorescence

1:10 - 1:500

Immunohistochemistry

1:1000-1:5000

Immunohistochemistry-Paraffin

1:10 - 1:500

Immunoprecipitation

1:10-1:500

Western Blot

1:3000-1:30000
Application Notes
This product is designed to detect enhanced GFP and GFP containing recombinant proteins. Tested in ELISA, IP, and WB and suitable in FACS, IHC, IF. This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen. Biotin conjugated monoclonal anti-GFP is well suited to titrate GFP in a sandwich ELISA in combination with polyclonal anti-GFP as the capture antibody. Only use the monoclonal form for the detection of enhanced or recombinant GFP. Polyclonal anti-GFP detects all variants of GFP tested to date. The biotin conjugated detection antibody is typically used with streptavidin conjugated HRP or other streptavidin conjugates. The use of polyclonal anti-GFP results in significant amplification of signal when fluorochrome conjugated polyclonal anti-GFP is used relative to the fluorescence of GFP alone. For immunoblotting use either alkaline phosphatase or peroxidase conjugated anti-GFP to detect GFP or GFP containing proteins on western blots. Optimal titers for applications should be determined by the researcher.

Use in In vitro assay reported in scientific literature (PMID 23125142).

Use in dot blot reported in scientific literature (PMID 24497578).

Use in immunoprecipitation reported in scientific literature (PMID 28198010).

Knockdown validation (Wolf et al).

Use in immunoblotting reported in scientific literature (PMID:31727786).

Reviewed Applications

Read 3 reviews rated 4.7 using NB600-597 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A 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. 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, clone 9F9.F9, GFP monoclonal

Additional GFP Products

Product Documents for GFP Antibody (9F9.F9) - BSA Free

Certificate of Analysis

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Product Specific Notices for GFP Antibody (9F9.F9) - 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 (9F9.F9) - BSA Free

Customer Reviews for GFP Antibody (9F9.F9) - BSA Free (3)

4.7 out of 5
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • GFP Antibody (9F9.F9)
    Name: Feng Li
    Application: Western Blot
    Sample Tested: 293 cell extract
    Species: Human
    Verified Customer | Posted 11/18/2019
    GFP fusion protein were overexpressed in 293T cells and can be detected by Nb600-597 antibody (Green). Specificity is good, but affinity is a little bit lower compared with Flag tag antibody.
    GFP Antibody (9F9.F9) - BSA Free NB600-597
  • GFP Antibody (9F9.F9)
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: HEK-293T cells transduced to have GFP
    Species: Human
    Verified Customer | Posted 05/10/2018
    The 293T cells have been transduced to have GFP which was detected by an antibody after permeabilization. The green staining shows the anti-GFP antibody.
    GFP Antibody (9F9.F9) - BSA Free NB600-597
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: CHOK1 cells transfected with GFP
    Species: Other
    Verified Customer | Posted 01/26/2015
    GFP NB600-597
    GFP Antibody (9F9.F9) - BSA Free NB600-597

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Protocols

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FAQs for GFP Antibody (9F9.F9) - BSA Free

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  • 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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