FLAG-Alpaca-iFluor488™ Anti-Flag tag Alpaca Nanobody (VHH), labelled iFluor™ 488

Overview

Description

FLAG-Alpaca-iFluor488™ Anti-Flag tag Alpaca Nanobody (VHH), labelled  iFluor™ 488

Reactivity

This antibody recognizes FLAG-tagged protein on Immunocytochemistry and Flow cytometry.

Tested applications

IF/ICC : 1:50-1:1000(dilution)

Flow Cytometry : Particularly suitable for a flow experiment

 

Because there is no Fc fragment of ordinary antibodies, there is no non-specific binding of Fc to the cell surface Fc receptor of immune cells in flow cytometry experiments

 

Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.

Product Picture

Product Picture 1

Figure 1. Alpaca antibodies are smaller than other mammalian antibodies. Mammalian IgG consists a heavy (green) and light (blue) chain, with variable heavy (VH) and variable light (VL) domains that facilitate the memory-like binding properties of antibodies. Camelid antibodies consist only a heavy chain (green) with a single variable heavy (VHH) domain for target recognition. This VHH domain can be expressed alone without the heavy chains, forming a single domain VHH.

 

Product Picture 2

Figure 2. Comparison of antibody types. Schematic representation of the comparison between (A) conventional antibodies, (B) alpaca heavy-chain antibodies, (C) and single domain antibodies.

Properties

Immunogen

Synthetic peptide DYKDDDDK conjugated to KLH

Clonality

Monoclonal nanobody, clone number: 3G11

Isotype

Alpaca VHH, Single Domain antibody

Form

In 10 mM phosphate buffered saline (PBS), pH 7.4, 50% glycerol with 1 mg/mL (0.1% w/v) BSA (IgG free, protease free) as a stabilizer and 0.02% (w/v) sodium azide as preservative.

Storage instruction

Store at -20°C for one year

Conjugation

iFluor™ 488
Excitation/Emission maxima wavelengths: 493 nm / 522 nm

Host

Applications

ICC/IF Image

Immunofluorescent analysis of (4% PFA) fixed HeLa cells using the iFluor 488-conjugated anti-FLAG nanobody (Green), at dilution of 1:200.

Application Image

Flow Cytometry experiment of HeLa cells using AT0066

 

An intracellular stain was performed on 1X106 HeLa cells with 1 ug FLAG nanobody (VHH, Single Domain antibody), labelled iFluor 488 (AT0066) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Both antibodies were conjugated to iFluor 488.

 

Highlights

Benefits Of Alpaca Nanobody

• No Fc fragment of ordinary antibodies, so there is no non-specific binding of Fc to the cell surface Fc receptor of immune cells in flow cytometry experiments
• Highly specific binding
• Low background
• High affinity
• High stability
• Alpaca nanobody is a small molecule with molecular weight of only 13kD (Conventional rabbit or mouse antibody is 150kD), so it can easily enter permeablized cells