Spot-Label for Western Blot

Details for Product No. ABIN5680072, Supplier: Log in to see
Target Name (Antigen)
Reactivity
Tag
Host
Camelid (Camelidae)
Antibody Type
Recombinant Antibody
Application
ELISA, Immunoprecipitation (IP), Western Blotting (WB)
Options
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Supplier Product No.
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Purpose Recombinant monovalent alpaca single-domain antibody, unconjugated, for the analysis of Spot-Tag fusion proteins.
Brand Spot-Tag®
Specificity Spot-Label binds to the Spot-Tag sequence PDRVRAVSHWSS both at the N-terminus or the C-terminus.
Characteristics Spot-Label is an anti-Spot-Nanobody (anti-Spot-VHH, Spot binding protein) either conjugated to fluorescent dyes or unconjugated. For Western blot (WB), the monovalent format of the Spot-Label is used: one anti-Spot Tag VHH/Nanobody labelled with fluorophores or unconjugated for detection by secondary antibody.
Purification Affinity-purified antibody fragment
Components Unconjugated Spot-Label for WB & IP, monovalent
Target Name (Antigen)
Target Type Tag
Background The Spot-Tag has an affinity-optimized 12 amino acid sequence to be strongly bind to the Spot-Nanobody. The inert tag can be bound to the N-terminal and to the C-terminal side of the protein of interest by PCR using modified primers or by using any of the Spot vectors offered. The expression of Spot-tagged proteins has been successfully tested in bacteria, yeast, mammalian cell lines, and insect cells so far.

Features:
- Small size
- PDRVRAVSHWSS
- 12 amino acid sequence
- 1.4 kDa
- pI: 10.4
- Hydrophilic
- Inert
- Linear
- Extinction coefficient: 5,500 L•mol-1•cm-1

Benefits:
- Easy to clone
- Works N-terminally and C-terminally
- Good expression
- Tested in bacteria, yeast, mammalian cell lines, and insect cells
Application Notes For Western blot (WB) and IP, the monovalent format of the Spot-Label (= Spot-Nanobody) is used: one anti-Spot-Nanobody connected labelled with fluorophores or unconjugated for detection by secondary antibody
Western blot: recommended starting dilution 1:500
ELISA: recommended starting dilution 1:1000 The optimal dilution depends on the application and should be determined by the end user.
For a secondary antibody, we recommend the use of anti-Llama IgG antibody, HRP-conjugated
Comment

The Spot-Tag® is a 12-amino acid epitope tag that is specifically recognized by monovalent Spot-Label, a 14.7 kDa small, high-affinity recombinant alpaca antibody fragment. The Spot-Tag system is useful for the purification of Spot-Tag fusion proteins and their analysis using immunoprecipitation, immunofluorescence, and super-resolution microscopy. Spot-Label can be used for Western blot, immunoprecipitation and ELISA analysis of Spot-Tag fusion proteins.

Restrictions For Research Use only
Format Liquid
Buffer Buffered aqueous solution, 0.09 % sodium azide
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze.
Storage 4 °C
Storage Comment refrigerate 4°C
Expiry Date 12 months
Supplier Images
Western Blotting (WB) image for Spot-Label for Western Blot (ABIN5680072) HEK293T cells spiked with Spot-tagged GFP Primary antibody: Spot-Label, monovalent 1:...
ELISA image for Spot-Label for Western Blot (ABIN5680072) Coating: mCherry-Spot and mCherry control (1 µg/well) Primary antibody: Spot-Label, m...
 image for Spot-Label for Western Blot (ABIN5680072) Schematics to visualize why size matters
 image for Spot-Label for Western Blot (ABIN5680072) Monovalent format of Spot-Label used for WB