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ChromoTek Nano-CaptureLigand™ mouse IgG2a, Fc-specific VHH, biotinylated

The Nano-CaptureLigand mouse IgG2a captures and immobilizes mouse IgG2a antibodies and Fc-fragments to avidin/streptavidin coated surfaces used in BLI, SPR, and ELISA assays. It consists of a biotinylated Nanobody that specifically binds to the mouse IgG2a subclass with high affinity and in a site-directed manner.

Applications

BLI, SPR, ELISA

Conjugate

Biotin

Host

Alpaca

Reactivity

Mouse IgG2b

Cat no : smsG2aB-1

Synonyms

Nano-CaptureLigand™ mouse IgG2a, Fc domain, mouse Ig2a, mouse immunoglobulin Gamma 2a

Validation Data Gallery

  • BLI binding kinetics of a mouse IgG2a anti-V5-tag antibody to a V5-tagged protein. A monoclonal mouse IgG2a anti-V5-tag antibody was immobilized using Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of a V5-tagged protein.

    BLI binding kinetics of a mouse IgG2a anti-V5-tag antibody to a V5-tagged protein. A monoclonal mouse IgG2a anti-V5-tag antibody was immobilized using Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of a V5-tagged protein.

  • Immobilization of the Nano-CaptureLigand (green) on Biosensor (white) for biolayer interferometry (BLI) followed by capture of mouse IgG2a antibody (orange). The setup is ready for characterization of mouse IgG2a antibodies’ binding kinetics and affinity to substrate (red), mouse IgG2a 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 mouse IgG2a antibody (orange). The setup is ready for characterization of mouse IgG2a antibodies’ binding kinetics and affinity to substrate (red), mouse IgG2a antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma.

  • ELISA capture of mouse IgG2a antibody using Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Mouse IgG2a antibody was titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

    ELISA capture of mouse IgG2a antibody using Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Mouse IgG2a 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 mouse IgG2a captures and immobilizes mouse IgG2a antibodies and Fc-fragments to avidin/streptavidin coated surfaces used in BLI, SPR, and ELISA assays. It consists of a biotinylated Nanobody that specifically binds to the mouse IgG2a subclass with high affinity and in a site-directed manner.

DescriptionThe next level of antibody immobilization to BLI and SPR biosensors:

• Gentle, site-directed and Fc-specific immobilization of mouse IgG2a antibodies and Fc-fragments 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 mouse IgG2a antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma samples

ApplicationsBLI, SPR, ELISA
HostAlpaca
ReactivityFc-fragment of mouse IgG2a
No cross-reactivity to mouse IgG1, Ig2b, IgG3; human IgG1, IgG2, IgG3, IgG4, IgA; rat IgG1, IgG2a, IgG2b; sera from goat, human, macaque (cynomolgus monkey), sheep
Cross-reactivity to mouse IgG2c
ConjugateBiotin
Physical StateLiquid
Purification MethodRecombinant expression, affinity purification IMAC
TypeNanobody
ClassRecombinant
Affinity (KD) KD=1.5 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_2848188
NoteThis product is for research use only, not for diagnostic or therapeutic use