Spot VHH
4
Description
Alpaca anti-Spot VHH, purified recombinant binding protein
Synonyms
Spot Nanobody, and Spot-Label (unconjugated protein)
More information about the Spot capture and detection tag can be found here.
Specificity
Spot- Binding Protein recognizes the Spot-tag sequence motif PDRVRAVSHWSS.
Applications
Coupling of dyes, biotin, etc. via NHS Ester reaction
Immunofluorescence (IF)
| Product | Size | Code | Price | Buy |
|---|---|---|---|---|
| Product Spot VHH, recombinant binding protein | Size 250 µL | Code etb-250 | Price $ 520 |
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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
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
RRID
AB_2827572
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 |
| V5 VHH | 14.6 | 22,710 | 8.0 |
| 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!