Spot VHH, recombinant binding protein

Description
Alpaca anti-Spot VHH, unconjugated, purified protein

Synonyms
Spot Nanobody, and Spot-Label (unconjugated protein)

Specificity
Spot- Binding Protein recognizes the Spot-tag sequence motif PDRVRAVSHWSS at the N-terminus or C-terminus of the Spot-tagged protein  

Applications
Coupling of dyes, biotin, etc. via NHS Ester reaction
Western blot, ELISA and IF

Product Size Code Price Buy
Product Spot VHH, recombinant binding protein Size 250 µL Code etb-250 Price $ 455
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Affinity
Dissociation constant KD of 6 nM

Specificity
Spot-Binding Protein recognizes the Spot-tag sequence motif PDRVRAVSHWSS at the N-terminus or C-terminus of the Spot-tagged protein

Conjugation:
Unconjugated
Suitable for conjugation of dyes, biotin, beads, surfaces etc. via NHS ester reaction
Spot VHH contains a sortase-tag and a His-tag at its C terminus: LPETGHHHHHH. This sequence includes the sortase recognition motif LPETG, for conjugation. In addition, bivalent Spot-VHH may be detected by an anti-His antibody. The Spot VHH corresponds to the “BC2-Nanobody” used by Virant et al in the publication "Nature Communications" (2018) doi:10.1038/s41467-018-03191-2.

Biophysical properties
Molecular weight MW: 30.3 kDa
Molecular extinction coefficient ε: 46,340 L·Mol−1·cm−1
Isoelectrical point pI: 5.3 (theoretical)

Form
Purified recombinant and bivalent antibody fragment.
Two anti-Spot-Tag VHHs/ Nanobodies genetically fused via a linker 30.4 kDa

Size
250 µL

Protein concentration
1 mg/mL

Storage buffer
25 mM TAPS pH8,5, 500 mM NaCl, 5 mM EDTA, preservative: 0.09 % sodium azide

Storage instructions
Shipped at ambient temperature. Upon receipt store at 4°C; stable for one year.

Do not freeze.

 

Are the nanobodies monoclonal or polyclonal?

The nanobodies are recombinant/monoclonal.

What are the biophysical parameters of the Nanobodies?

Name MW (kDa) Molar Extinction Coefficient
(M-1 cm-1)
Theoretical pI
GFP VHH 13.9 27.055 9.4
GST VHH 14.8 28.545 8.2
Halo VHH 14.8 23.045 9.4
Histon VHH 14.4 25.565 10.2
MBP VHH 14.1 27.055 10.0
Mdm4/Hdmx VHH 13.6 14.565 10.1
MK2 VHH 14.4 32.555 9.7

mNeonGreen VHH

14.3 21.680 8.0
Myc VHH 13.5 23.045 9.6
p53 C-term VHH 14.8 34.170 9.2
p53 N-term VHH 13.0 20.065 9.7
PARP1 VHH 14.4 17.670 9.2
RFP VHH 14.9 30.035 9.3
SNAP/CLIP-tag VHH 13.1 24.075 9.8
Spot VHH (bivalent) 30.3 46.340 5.3
TurboGFP VHH 13.6 18.575 9.5
Vimentin VHH 26.7 48.150 10.0

Do the nanobodies have any tags?

Yes, the nanobodies have a C-terminal His6-tag.

Extraordinary stable & reliable binding

  • Stable up to 65 °C, 10 mM DTT, 0.75 M Guanidinium•HCl, 2 M Urea, 2 M NaCl, 2 % Nonidet P40 Substitute, 0.1 % SDS, 1 % Triton X-100, 1 % Dexoycholate
  • pH 4-10
  • Fulfills highest requirements for antibody validation
  • Structure and function are characterized

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Only for research applications, not for diagnostic or therapeutic use!