ChromoTek Spot-Label® ATTO594

Validation Data Gallery

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  STED: IF of Spot-tagged Actin-Chromobody with Spot-Label Atto594 bivalent (1:1,000). Gated STED images were acquired with a Leica TCS SP8 STED 3X microscope with pulsed White Light Laser excitation at 590 nm and pulsed depletion with a 775 nm laser. Objective: 100x Oil STED White, NA: 1.4. Pixel size: 21 x 21 nm; z-Step size of z-Stacks: 0.16 μm. Images were deconvolved with Huygens Professional (SVI). STED images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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  Application of Spot-tag related products.

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  Widefield images of HeLa cells expressing Tom70-EGFP-Spot-Tag immunostained with Spot-Label ATTO594, bivalent (1:5,000). Left: GFP channel, middle: channel for ATTO594, right: overlay, also with DAPI. Scale bar, 10 μm.

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  Confocal images of HeLa cells expressing Tom70-EGFP-Spot-Tag immunostained with Spot-Label ATTO594 bivalent (1:1,000). Left: EGFP fluorescence, middle: ATTO594 signal, right: merge. Scale bar, 10 μm. The images were acquired with Leica TCS SP8 microscope, 100X oil objective, at the Core Facility Bioimaging at the Biomedical Center, LMU Munich, and deconvolved with Huygens Professional (SVI). Left: GFP fluorescence, middle: ATTO594 signal, right: merge. Scale bar, 10 μm.

anti-Spot-Tag® VHH/ Nanobody conjugated to fluorescent dye for immunofluorescence, microscopy, and immunoblotting of Spot-tagged proteins

Cat No. eba594

Publications(3)

Specificity

Spot-Tag^®

Applications

IF

Conjugate

ATTO 594

Host

Alpaca

Synonyms

Spot tag, Spot, PDRVRAVSHWSS, Spot-Label

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Product Information

anti-Spot-Tag® VHH/ Nanobody conjugated to fluorescent dye for immunofluorescence, microscopy, and immunoblotting of Spot-tagged proteins

• Description: Immunofluorescence of Spot-tagged proteins with anti-Spot Nanobody conjugated to fluorescent dye. Spot-Label is much smaller compared to conventional primary plus secondary IgG antibodies (IgG complex). Due to its small size, Spot-Label is optimal for effective labeling with minimal fluorophore displacement for super-resolution microscopy.

  • Capture and detection tag without compromises in applications

  • Higher image resolution

  • Superior accessibility and labelling of epitopes in crowded cellular/organelle environments

  • Strong avidity effect from bivalent form of Spot-VHH

  • Fulfills highest requirements on antibody validation: structure and function characterized

• Applications: IF
• Host: Alpaca
• Specificity/Target: Spot-Tag^® (PDRVRAVSHWSS)
• Conjugate: ATTO 594
• Physical State: Liquid
• Suggested Dilution: IF: 1:1,000
• Type: Nanobody
• Class: Recombinant
• RRID: AB_2827570
• Affinity (KD): Dissociation constant K_D of 6 nM
• Spot-Tag® Origin: Engineered 12 amino acid sequence modified from the unstructured N-terminus of beta-catenin
• Cross Reactivity: The Spot-Nanobody shows minimal cross reactivity with beta-catenin.
• Excitation/ Emission: Excitation range 580 - 615 nm (λ_abs= 601 nm) ; Emission range 620 - 660 nm (λ_fl= 627 nm)
• Storage Buffer: 1x PBS, 0.09% sodium azide
• Storage Condition: Shipped at ambient temperature. Upon receipt store at +4°C. Stable for 6 month. Do not freeze. Protect from light.
• Size: 10 μL; 50 μL
• Note: This product is for research use only, not for diagnostic or therapeutic use

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Publications

Application	Title
IF	Microbiol Spectr; BopN is a Gatekeeper of the Bordetella Type III Secretion System; Authors - Kevin Munoz Navarrete
IF	Methods Mol Biol; Superresolution Expansion Microscopy in Dictyostelium Amoebae; Authors - Marianne Grafe
IF	Cell Death Differ; Inhibition of BAK-mediated apoptosis by the BH3-only protein BNIP5; Authors - Sebastian Rühl