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ChromoTek Nano-CaptureLigand® human IgG/rabbit IgG, Fc-specific VHH, biotinylated

The Nano-CaptureLigand® human IgG captures and immobilizes the Fc-fragment of human IgG antibodies to avidin/streptavidin coated surfaces used in BLI, SPR, and ELISA assays. It consists of a biotinylated Nanobody that specifically binds to human IgG antibodies with high affinity and in a site-directed manner.

Applications

BLI, SPR, ELISA

Conjugate

Biotin

Host

Alpaca

Reactivity

Human, Rabbit

Cat No : shurbGB-1

Synonyms

immunoglobulin, human IgG, Fc domain, Rabbit IgG

Validation Data Gallery

  • BLI binding kinetics of a monoclonal rabbit IgG anti-DYKDDDDK-tag antibody to a DYKDDDDK-tagged protein. A monoclonal rabbit IgG anti-DYKDDDDK-tag antibody was immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of a DYKDDDDK-tagged protein.

    BLI binding kinetics of a monoclonal rabbit IgG anti-DYKDDDDK-tag antibody to a DYKDDDDK-tagged protein. A monoclonal rabbit IgG anti-DYKDDDDK-tag antibody was immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of a DYKDDDDK-tagged protein.

  • ELISA capture of a rabbit IgG antibody using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Rabbit IgG antibody were titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

    ELISA capture of a rabbit IgG antibody using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Rabbit IgG antibody were titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

  • BLI binding kinetics of a polyclonal rabbit IgG anti-GFP antibody to eGFP. A polyclonal rabbit IgG anti-GFP antibody (ChromoTek GFP antibody [PABG1]) was immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of eGFP (ChromoTek EGFP, recombinant protein).

    BLI binding kinetics of a polyclonal rabbit IgG anti-GFP antibody to eGFP. A polyclonal rabbit IgG anti-GFP antibody (ChromoTek GFP antibody [PABG1]) was immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of eGFP (ChromoTek EGFP, recombinant protein).

  • BLI binding kinetics of different human IgG antibodies. Different monoclonal human IgG antibodies were immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors. All four human IgG subclasses are stably captured by the Nano-CaptureLigand with negligible dissociation.

    BLI binding kinetics of different human IgG antibodies. Different monoclonal human IgG antibodies were immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors. All four human IgG subclasses are stably captured by the Nano-CaptureLigand with negligible dissociation.

  • Immobilization of the Nano-CaptureLigand (green) on Biosensor (white) for biolayer interferometry (BLI) followed by capture of human IgG antibody (orange). The setup is ready for characterization of human IgG antibodies’ binding kinetics and affinity to substrate (red), human IgG antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma.

    Immobilization of the Nano-CaptureLigand (green) on Biosensor (white) for biolayer interferometry (BLI) followed by capture of human IgG antibody (orange). The setup is ready for characterization of human IgG antibodies’ binding kinetics and affinity to substrate (red), human IgG antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma.

  • ELISA capture of human IgG1 lambda antibody using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Human IgG1 lambda antibody was titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

    ELISA capture of human IgG1 lambda antibody using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Human IgG1 lambda antibody was titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

  • Sandwich ELISA: Immobilization of the Nano-CaptureLigand on streptavidin coated microplates for ELISA. No biotinylation of the antibody is required. Coating of the streptavidin microplate with Nano-CaptureLigand (top left). Site-directed antibody immobilization via Nano-CaptureLigand (top right). Addition of sample including target (red, bottom left). Addition of detection and secondary antibody for read-out (bottom right).

    Sandwich ELISA: Immobilization of the Nano-CaptureLigand on streptavidin coated microplates for ELISA. No biotinylation of the antibody is required. Coating of the streptavidin microplate with Nano-CaptureLigand (top left). Site-directed antibody immobilization via Nano-CaptureLigand (top right). Addition of sample including target (red, bottom left). Addition of detection and secondary antibody for read-out (bottom right).

  • Species reactivity of Secondary VHH.

    Species reactivity of Secondary VHH.

Product Information

The Nano-CaptureLigand® human IgG captures and immobilizes the Fc-fragment of human IgG antibodies to avidin/streptavidin coated surfaces used in BLI, SPR, and ELISA assays. It consists of a biotinylated Nanobody that specifically binds to human IgG antibodies with high affinity and in a site-directed manner.

DescriptionThe next level of antibody immobilization to BLI and SPR biosensors

• Site-directed and specific immobilization of the Fc-fragment of human IgG and rabbit IgG to streptavidin coated biosensors without biotinylation of the antibody

• Compatible to surface plasmon resonance (SPR) and biolayer interferometry (BLI) assays (e.g. FortéBio Octet® systems)

• High affinity for stable baseline without significant dissociation

• >10x regeneration for multiple reuse

• High specificity enables Fc-fragment of human IgG and rabbit IgG antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma samples

ApplicationsBLI, SPR, ELISA
HostAlpaca
ReactivityHuman IgG1, IgG2, IgG3, and IgG4 Fc-fragments
Rabbit IgG Fc-fragment
ConjugateBiotin
Physical StateLiquid
Purification MethodRecombinant expression, affinity purification IMAC
TypeNanobody
ClassRecombinant
Affinity (KD) KD:Human IgG1: 1.4 nM; Human IgG2: 0.4 nM; Human IgG3: 9.0 nM; Human IgG4: 0.2 nM
Rabbit IgG: 0.2 nM
Apparent affinity may be higher for full IgGs due to avidity effects (1 antibody captured by 2 Nanobodies).
Storage Buffer25 mM TAPS pH 8.5, 500 mM NaCl, 5 mM EDTA, Preservative: 0.09 % sodium azide
Storage ConditionShipped at ambient temperature. Upon receipt store at +4°C/+40°F. Optional: Aliquot upon arrival and store at -20°C/-4°F
RRIDAB_2848185
NoteThis product is for research use only, not for diagnostic or therapeutic use