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  1. Link

    gfp-antibody-ab13970.pdf

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Tags & Cell Markers Fusion / Marker Proteins GFP
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Anti-GFP antibody (ab13970)

  • Datasheet
  • SDS
Reviews (92)Q&A (21)References (2684)

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Western blot - Anti-GFP antibody (ab13970)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
  • Western blot - Anti-GFP antibody (ab13970)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
  • Western blot - Anti-GFP antibody (ab13970)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)

Key features and details

  • Chicken polyclonal to GFP
  • Suitable for: WB, ICC/IF
  • Reacts with: Species independent
  • Isotype: IgY

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-GFP antibody [EPR14104] (ab183734)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-GFP antibody
    See all GFP primary antibodies
  • Description

    Chicken polyclonal to GFP
  • Host species

    Chicken
  • Specificity

    Our GFP antibody does cross-react with the many fluorescent proteins that are derived from the jellyfish Aequorea victoria. These are all proteins that differ from the original GFP by just a few point mutations (EGFP, YFP, mVenus, CFP, BFP etc.).

  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Recombinant full length protein corresponding to GFP.
    Database link: P42212

  • Positive control

    • ICC: GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line). WB: Transgenic mouse spinal cords.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7
    Preservative: 0.01% Thimerosal (merthiolate)
    Constituents: PBS, 50% Glycerol, 0.16% Sodium phosphate
  • Concentration information loading...
  • Purity

    IgY fraction
  • Purification notes

    Sterile filtered.
  • Clonality

    Polyclonal
  • Isotype

    IgY
  • Research areas

    • Tags & Cell Markers
    • Fusion / Marker Proteins
    • GFP
    • Signal Transduction
    • Antibodies
    • gfp

Associated products

  • Compatible Secondaries

    • Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173)
  • Isotype control

    • Chicken IgY - Isotype Control (ab50579)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • Mouse lung normal tissue lysate - total protein (ab29297)
    • Mouse brain tissue lysate - total protein (ab30151)
  • Recombinant Protein

    • Recombinant E. coli GFP protein (His tag) (ab119740)
    • Recombinant A. victoria GFP protein (ab84191)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Recombinant Enhanced GFP protein (His tag) (ab134853)
    • GFP ELISA Kit (ab171581)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab13970 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
WB (13)
1/5000.
ICC/IF (22)
1/2000.

Used at a dilution of 1/2000 for 1 hr (see Abreview for further information).

Notes
WB
1/5000.
ICC/IF
1/2000.

Used at a dilution of 1/2000 for 1 hr (see Abreview for further information).

Target

  • 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

Images

  • Western blot - Anti-GFP antibody (ab13970)
    Western blot - Anti-GFP antibody (ab13970)This image is courtesy of an anonymous Abreview
    All lanes : Anti-GFP antibody (ab13970) at 1/2000 dilution (Diluent 1x TBS /4 hours at 4°C)

    Lane 1 : 3 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature
    Lane 2 : 2 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature
    Lane 3 : 1 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature

    Lysates/proteins at 25 µg per lane.

    Blocking peptides at 5 % per lane.

    Secondary
    All lanes : Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (ab150176) at 1/5000 dilution

    Performed under reducing conditions.

    Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds


    Gel Running Conditions: Reduced Denaturing (15% PAGE)

    Detection method: Fluorescent Secondary Antibodies

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)This image is courtesy of an Abreview submitted by Christophe Lachaud

    ab13970 staining GFP in U-2 OS (Human bone osteosarcoma epithelial cell line) cells by ICC/IF.

    Cells were paraformaldehyde fixed, permeabilized with 0.5% triton and blocked with 2% antibody dilution buffer for 2 hours. Cells were incubated with the primary antibody (1/1000) for 1 hour at 25°C. An undiluted Alexa Fluor® 488 conjugated Goat anti-chicken polyclonal was used as the secondary antibody.

    See Abreview

  • Western blot - Anti-GFP antibody (ab13970)
    Western blot - Anti-GFP antibody (ab13970)Image courtesy of an anonymous Abreview.
    All lanes : Anti-GFP antibody (ab13970) at 1/1000 dilution

    Lane 1 : Whole cell lysate prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
    Lane 2 : Whole cell lysate prepared from HeLa cells.

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : IRDye 800CW conjugated goat anti-chicken polyclonal at 1/15000 dilution

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)

    Transgenic mice expressing GFP selectively in lamina II of the spinal cord.

    In the right panels, note the correspondance between the green (rabbit anti-GFP) and red signals (chicken anti-GFP from Abcam) indicating that these two antibody preparations recognized the same gene product. The secondary antibody used with ab13970 was an FITC-labeled goat anti-chicken

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)

    ab13970 staining GFP in GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line) cells.

    The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab13970 at 1/2000 dilution overnight at +4°C followed by incubation with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173), for 1 hour, at 1 μg/ml.
    Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH/3T3 cells, the cells upon which ab13970 was applied gave a stronger signal in the 488 channel, indicating that ab13970 is binding to GFP and therefore eliciting signal amplification.
    ab13970 was also applied to non-GFP-transfected NIH/3T3 cells, which produced no positive staining, indicating specificity for GFP.

    Nuclear DNA was labeled with 1.43 μM DAPI (blue).

  • Western blot - Anti-GFP antibody (ab13970)
    Western blot - Anti-GFP antibody (ab13970)
    Lane 1 : Rabbit anti-GFP
    Lane 2 : Anti-GFP antibody (ab13970)

    All lanes : Transgenic mouse spinal cords


    Western blot of transgenic mouse spinal cords showing that the rabbit anti-GFP (lane 1) and the chicken anti-GFP (Abcam; lane 2) recognize a band at the same molecular weight.

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)This image is courtesy of an Abreview submitted by Dr Radbod Darabi

    ab13970 staining GFP + tumor in mouse muscle cells by ICC/IF.

    Cells were formaldehyde fixed and blocked with 3% BSA for 1 hour at 24°C prior to incubation with the primary antibody (1/500) for 1 hour at 24°C. An Alexa Fluor® 488 conjugated goat anti-chicken was used as the secondary.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)Image courtesy of an AbReview submitted by Dr Francois Daubeuf.

    Immunocytochemical/ Immunoflurescence analysis of cytospined HEK-293 cells (Human epithelial cell line from embryonic kidney) transfected with GFP, labeling GFP with ab13970 at 1/200 incubated for 16 hours at 4°C with 1% BSA in PBS. Secondary used was a donkey anti-chicken polyclonal DyLight® 594 at 1/500.

    GFP is shown in red (DyLight® 594).

    Nuclei are counterstained in blue (DAPI).

    The left panel shows HEK-293 cells transfected with GFP and the right panel shows non-transfected HEK-293 cells.

    See Abreview

Protocols

  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2684)

Publishing research using ab13970? Please let us know so that we can cite the reference in this datasheet.

ab13970 has been referenced in 2684 publications.

  • Horita N  et al. Delta-like 1-Expressing Cells at the Gland Base Promote Proliferation of Gastric Antral Stem Cells in Mouse. Cell Mol Gastroenterol Hepatol 13:275-287 (2022). PubMed: 34438113
  • La Rocca G  et al. Inducible and reversible inhibition of miRNA-mediated gene repression in vivo. Elife 10:N/A (2021). PubMed: 34463618
  • Hardcastle BJ  et al. A visual pathway for skylight polarization processing in Drosophila. Elife 10:N/A (2021). PubMed: 33755020
  • Feng S  et al. Scarless engineering of the Drosophila genome near any site-specific integration site. Genetics 217:N/A (2021). PubMed: 33772309
  • Schick E  et al. Early cis-regulatory events in the formation of retinal horizontal cells. Dev Biol 476:88-100 (2021). PubMed: 33774011
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 113 Abreviews or Q&A

IHC - Wholemount abreview for Anti-GFP antibody

Excellent
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Fruit fly (Drosophila melanogaster) Tissue (Adult Posterior Midgut, HEAT-FIXED)
Specification
Adult Posterior Midgut, HEAT-FIXED
Read More

Dr. Joaquin Denavascues

Verified customer

Submitted Feb 12 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (HEK293)
Specification
HEK293
Permeabilization
Yes - 0.2% triton
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Sep 26 2013

Western blot abreview for Anti-GFP antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Non-reduced Denaturing
Sample
Human Cell lysate - whole cell (HEK 293)
Specification
HEK 293
Treatment
transfection with 1 ug GFP plasmid
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Aug 01 2013

Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Abdil buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2%
Sample
Human Cell (U2OS)
Specification
U2OS
Permeabilization
Yes - 0.5% triton
Fixative
Paraformaldehyde
Read More

DR. Christophe Lachaud

Verified customer

Submitted Aug 01 2013

Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (U2OS cells)
Specification
U2OS cells
Permeabilization
Yes - 0.2% Triton
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jul 26 2013

Immunohistochemistry free floating abreview for Anti-GFP antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (brain)
Specification
brain
Read More

Mr. Binaree .

Verified customer

Submitted Nov 16 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GFP antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mouse brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate buffer
Permeabilization
Yes - 0.2% TritonX-100 in PBS for 3hours along with blocking solution.
Specification
mouse brain
Blocking step
Serum as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 28°C
Fixative
Paraformaldehyde
Read More

MR. Lenin Veeraval

Verified customer

Submitted Aug 12 2021

Immunocytochemistry abreview for Anti-GFP antibody

Average
Abreviews
Abreviews
Application
Immunocytochemistry
Sample
Mouse Cell (heart)
Permeabilization
No
Specification
heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Apr 15 2021

Immunocytochemistry abreview for Anti-GFP antibody

Average
Abreviews
Abreviews
Application
Immunocytochemistry
Sample
Rat Cell (heart)
Permeabilization
No
Specification
heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 37°C
Fixative
Formaldehyde
Read More

Ann Kurian

Verified customer

Submitted Mar 30 2021

Question

Can you please let me know what positive control can I use for Anti-GFP antibody (ab13970)

Read More

Abcam community

Verified customer

Asked on Aug 17 2012

Answer

I would recommend the following GFP protein to use as a positive control on a Western Blot:

https://www.abcam.com/GFP-protein-ab84191.html.


Read More

Abcam Scientific Support

Answered on Aug 17 2012

1-10 of 113 Abreviews or Q&A

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