Overview

  • Product name
  • Description
    Goat polyclonal to GFP
  • Host species
    Goat
  • Specificity
    Reactive against all variants of Aequorea victoria GFP such as S65T-GFP, RS-GFP, YFP and EGFP.
  • Tested applications
    Suitable for: WB, IPmore details
  • Immunogen

    This information is considered to be commercially sensitive.

  • General notes
    Protein A will not bind goat IgG, so use alternates (eg. protein G) in IP with this antibody. This antibody is available in an affinity purified form as ab5450.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.05% Sodium azide
  • Purity
    Whole antiserum
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5449 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 1/5000 - 1/20000.
IP Use 0.5µl for 106 cells.

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

  • Lane 1       :  parental YTS cells (negative control)
    Lanes 2-5 :  YTS cells transfected with KIR-EGFP (mw 88 kD)

    KIR-EGFP IP's with Goat polyclonal to GFP (ab5449) using 0.1 ul for 2x106 cells. KIR-EGFP detected with a mouse monoclonal to KIR receptor (Borszcz et al EGI 2003, 33: 1084).

     



    Lane 1 : parental YTS cells (negative control) Lanes 2-5 : YTS cells transfected with KIR-EGFP (mw 88 kD) KIR-EGFP IP's with Goat polyclonal to GFP (ab5449) using 0.1 ul for 2x106 cells. KIR-EGFP detected with a mouse monoclonal to KIR receptor (Borszcz et al EGI 2003, 33: 1084).
  • 5 ng GFP on  PVDF membrane QC. Goat polyclonal to GFP (ab5449) used at dilutions of:
    Lane 1 : 1/2500
    Lane 2 : 1/5000
    Lane 3 :  1/10,000
    Lane 4 :  1/20,000



    5 ng GFP on PVDF membrane QC. Goat polyclonal to GFP (ab5449) used at dilutions of: Lane 1 : 1/2500 Lane 2 : 1/5000 Lane 3 : 1/10,000 Lane 4 : 1/20,000

References

This product has been referenced in:
  • Kim MY  et al. Mbd2-CP2c loop drives adult-type globin gene expression and definitive erythropoiesis. Nucleic Acids Res 46:4933-4949 (2018). Read more (PubMed: 29547954) »
  • Boubriak II  et al. Stress-induced release of Oct-1 from the nuclear envelope is mediated by JNK phosphorylation of lamin B1. PLoS One 12:e0177990 (2017). Read more (PubMed: 28542436) »
See all 6 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (Hansenula polymorpha)
Loading amount
10 µg
Specification
Hansenula polymorpha
Gel Running Conditions
Reduced Denaturing (12)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 28 2009

Answer

Thank you for your enquiry. I can confirm that ab5450 (affinity purified) and ab5449 (crude) are exactly what you are looking for. These antibodies were made in goat using the exact same antigen and they work just as well as ab6556 in all applications tested.

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Answer

Thank you for your enquiry. If your CFP is derived from the GFP of Aequorea victoria jellyfish, then I am afraid all our anti-GFP antibodies will cross-react with that protein, since the difference is a single amino-acid substitution and none of our anti-GFP antibodies are directed against that epitope. If your CFP is from another species, for example the reef coral Anemonia majano, then the antibodies will not cross-react. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your interesting enquiry. I have had a look at the details for the immunogens on these antibodies and obtained some details for you. ab1218 is a mouse monoclonal which is been raised against a recombinant GFP fusion protein using amino acids 1-246 of GFP from jellyfish Aiquorea Victoria. It is reactive against all varients of this protein. Ab5449 is a goat polyclonal to the full length recombinant GFP protein (from Aiquorea Victoria) prepared in e coli. Its concentration is not available as this antibody is supplied as whole goat antiserum. Ab5450 is the same as Ab5449 but it has been affinipured. This means it is likely to be more specific and stronger binding to the immunogen. Its concentration is 0.5mg/ml.

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Question
Answer

Yes, it is very likely it will detect EGFP as well.

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