Recombinant

Recombinant Anti-GFP VHH Single Domain antibody (Sepharose) (ab191863)

Overview

  • Product name

    Anti-GFP VHH Single Domain antibody (Sepharose)
    See all GFP VHH Single Domain primary antibodies
  • Description

    Llama monoclonal to GFP VHH Single Domain (Sepharose)
  • Host species

    Llama
  • Conjugation

    Sepharose
  • Specificity

    Anti-GFP VHH Single Domain Antibody (Sepharose) (ab191863) specifically binds GFP, eGFP and BFP. ab191863 binds GFP N-terminal and C-terminal fusion proteins. It does not interact with CFP, RFP, YFP, mCherry or TurboGFP.
  • Tested applications

    Suitable for: IPmore details
  • Immunogen

    Recombinant full length protein corresponding to Aequorea victoria GFP VHH Single Domain.

  • Positive control

    • This antibody gave a positive signal in NIH3T3 overexpressing GFP cell line within IP.
  • General notes

    Sepharose® is a registered trademark of GE Healthcare.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at 4°C (stable for up to 12 months). Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.40
    Preservative: 0.05% Sodium azide
    Constituents: PBS, Sodium phosphate, Sodium chloride
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    This product is a recombinant protein produced in E. coli.
  • Clonality

    Monoclonal
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab191863 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
AP Use at an assay dependent concentration.

20µl of slurry pulls down a minimum of 5-10 µg GFP

IP Use at an assay dependent concentration.

20µl of slurry pulls down a minimum of 5-10 µg GFP

Target

  • Relevance

    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. 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
    • yfp antibody

Images

  • Lane 1: Anti-GFP VHH Single Domain Antibody (Sepharose) (ab191863) + 10ul recombinant GFP (ab84191)

    Lane 2: Anti-GFP VHH Single Domain Antibody (Sepharose) (ab191863) +10ul GFP fusion protein

    Lane 3: Off-target sdAb beads (Anti-Rabbit VHH Single Domain Antibody (Sepharose) (ab191867) + 10ul recombinant GFP (ab84191)

    Lane 4: Anti-GFP antibody (ab290) + Protein A agarose + 10ul recombinant GFP (ab84191)

    Lane 5: Anti-GFP antibody (ab290) + Protein A agarose + 10ul GFP fusion protein

     

    Anti-GFP VHH Single Domain Antibody Sepharose (ab191863) beads were used to pull down recombinant GFP (ab84191) and GFP fusion protein. ab290 and Protein A agarose were used as a positive control. The gel was stained with Optiblot Blue (ab119211).

  • Lane 1: NIH-3T3 cell lysate overexpressing GFP - 10ug

    Lane 2: Unbound IP supernatant: Anti-GFP VHH Single Domain Antibody (Sepharose) (ab191863) + 100ug NIH-3T3 cell lysate overexpressing GFP

    Lane 3: Negative IP: off-target sdAb beads (Anti-Rabbit VHH Single Domain Antibody (Sepharose) (ab191867) + 100ug NIH-3T3 cell lysate overexpressing GFP

    Lane 4: IP: Anti-GFP VHH Single Domain Antibody (Sepharose) (ab191863) + 100ug NIH-3T3 cell lysate overexpressing GFP

    Lane 5: NIH-3T3 cell lysate overexpressing GFP - 10ug

    Lane 6: Unbound IP supernatant: anti-GFP antibody (ab290) + Protein A agarose + 100ug NIH-3T3 cell lysate overexpressing GFP

    Lane 7: IP: anti-GFP antibody (ab290) + Protein A agarose+ 100ug NIH-3T3 cell lysate overexpressing GFP

     

    Anti-GFP VHH Single Domain Antibody (Sepharose) ab191863 beads were used to pull down GFP from NIH-3T3 cell lysate overexpressing GFP. ab290 and Protein A agarose were used as a positive control. Note the light chain of IgG obscuring GFP on Lane 7. The gel was stained with Optiblot Blue (ab119211).



    All lanes :

    Lanes 1 & 5 : NIH-3T3 cell lysate overexpressing GFP - 10ug
    Lanes 2 & 6 : NIH-3T3 cell lysate overexpressing GFP - IP supernatant - 10ug
    Lane 3 : Off target sdAb beads pull down
    Lane 4 : ab191863 beads pulled down GFP protein (27kDa)
    Lane 7 : Sample: ab290 beads pulled down GFP protein (27kDa), obscurred by light chain

    Observed band size: 27 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 25 kDa (possible IgG), 50 kDa (possible IgG)

  • Lane 1: NIH-3T3 cell lysate overexpressing GFP - 10ug

    Lane 2: Unbound IP supernatant: Anti-GFP VHH Single Domain Antibody (Sepharose) (ab191863) + 100ug NIH-3T3 cell lysate overexpressing GFP

    Lane 3: IP: Anti-GFP VHH Single Domain Antibody (Sepharose) (ab191863) + 100ug NIH-3T3 cell lysate overexpressing GFP

    Lane 4: Negative IP: off-target sdAb beads (Anti-Rabbit VHH Single Domain Antibody (Sepharose) (ab191867) + 100ug NIH-3T3 cell lysate overexpressing GFP

    Lane 5: NIH-3T3 cell lysate overexpressing GFP - 10ug

    Lane 6: Unbound IP supernatant: anti-GFP antibody (ab290) + Protein A agarose + 100ug NIH-3T3 cell lysate overexpressing GFP

    Lane 7: IP: anti-GFP antibody (ab290) + Protein A agarose+ 100ug NIH-3T3 cell lysate overexpressing GFP

     

    Anti-GFP VHH Single Domain Antibody (Sepharose) (ab191863) beads were used to pull down GFP from NIH-3T3 cell lysate overexpressing GFP. ab290 and Protein A agarose were used as a positive control. The gel was transferred to Nitrocellulose membrane and probed with mouse anti-GFP (ab1218) and anti-mouse HRP (ab97040).



    All lanes :

    Lanes 1 & 5 : NIH-3T3 cell lysate overexpressing GFP - 10ug
    Lanes 2 & 6 : NIH-3T3 cell lysate overexpressing GFP - IP supernatant - 10ug
    Lane 3 : ab191863 beads pulled down GFP protein (27kDa)
    Lane 4 : Off target sdAb beads pull down

References

ab191863 has not yet been referenced specifically in any publications.

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