PFA fixed COS cells transfected with mScarlet-i-tubulin (A) stained with FluoTag-X2 anti-mScarlet-i Atto488 (B). Overlay in (C).

FluoTag-X2 anti-mScarlet-i At488 (mScarlet3)

Immunofluorescence analysis of KOLF2.1J human iPSCs transfected with pmScarlet3_C1 plasmid (Addgene #189753). Post-fixation, cells were stained with FluoTag®-X2 anti-mScarlet-i coupled to Atto 488 (Cat. No. N1302-At488, dilution 1:500). A The N1302-At488 specific signal is shown in green. B The mScarlet3 fluorescence is shown in red. C Nuclei were visualized by DAPI staining. D Merge of A – C confirms that FluoTag®-X2 anti-mScarlet-i specifically recognizes the mScarlet3 variant. The images were captured under 40x magnification. Image courtesy of Victoria Menne, King’s College London.

FluoTag-X2 anti-mScarlet-i AF647

PFA-fixed Cos7 cells expressing a TOM70-nfmScarlet fusion protein (nf: non-fluorescent) were stained with FluoTag®-X2 anti-mScarlet-i coupled to Alexa Fluor 647 (Cat. No. N1302-AF647, dilution 1:500). A Greyscale image of the staining performed with N1302-AF647. B False color representation of the image shown in A is displayed in magenta (coloring according to the excitation wavelength of the employed fluorophore). C The corresponding DAPI signal of the depicted section. D Merge of A and C. False color representation of A in magenta and C in blue.

FluoTag-X2 anti-mScarlet-i AF568

PFA-fixed Cos7 cells expressing a TOM70-nfmScarlet fusion protein (nf: non-fluorescent) were stained with FluoTag®-X2 anti-mScarlet-i coupled to AZDye 568 (Cat. No. N1302-AF568, dilution 1:500). A Greyscale image of the staining performed with N1302-AF568. B False color representation of the image shown in A is displayed in red (coloring according to the excitation wavelength of the employed fluorophore). C The corresponding DAPI signal of the depicted section. D Merge of A and C. False color representation of A in red and C in blue.

FluoTag-X2 anti-mScarlet-i At488

PFA-fixed Cos7 cells expressing a TOM70-nfmScarlet fusion protein (nf: non-fluorescent) were stained with FluoTag®-X2 anti-mScarlet-i coupled to Atto 488 (Cat. No. N1302-At488, dilution 1:500). A Greyscale image of the staining performed with N1302-At488. B False color representation of the image shown in A is displayed in green (coloring according to the excitation wavelength of the employed fluorophore). C The corresponding DAPI signal of the depicted section. D Merge of A and C. False color representation of A in green and C in blue.

FluoTag®-X2 anti-mScarlet-i

Cat No: N1302 Category:

400,00 

FluoTag®-X2 anti-mScarlet is derived from our sdAb anti-RFP (clone 2B12) and binds specifically and strongly to many mScarlet variants (validated for mScarlet-I, mScarlet-H and mScarlet-3). It also recognizes DsRed, mRFP, mCherry and others DsRed family members. Conjugated to fluorophores, it enables direct and highly specific fluorescent detection, eliminating the need for secondary antibodies.

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mScarlet was created in vitro using a synthetic consensus template from various red fluorescent proteins (ref). It is one of the brightest (~3x brighter than mCherry) and truly monomeric fluorescent proteins, making it ideal for cellular imaging (FPbase).  mScarlet can generally be employed in two distinct versions. Each version carries a distinct single amino acid substitution, mScarlet-I (T74I) which features an accelerated maturation speed, and mScarlet-H (M164H) which displays higher photostability. An open-source database with the most currently available variants can be found in the fluorescence protein database “fpbase”.

FluoTag®-X2 anti-mScarlet-i is derived from our sdAb anti-RFP (clone 2B12) and binds specifically and strongly to many mScarlet variants (validated for mScarlet-I, mScarlet-H and mScarlet-3). It also recognizes DsRed, mRFP, mCherry and others DsRed family members. For a detailed overview, refer to our FP Specificity Chart.

FluoTag®-X2 anti-mScarlet-i is directly conjugated to two fluorophores per sdAb (FluoTag®X2 variant). To learn more about the FluoTags®, please visit our Technology section here.

Variations:
Conjugation Amount Cat No. RRID
Atto488 200 μl N1302-At488-L AB_3075969
AZDye568 200 μl N1302-AF568-L AB_3075967
Atto643 200 μl N1302-At643-L AB_3075970
Alexa647 200 μl N1302-AF647-L AB_3075968
AbberiorStar635P 200 μl N1302-Ab635P-L AB_3075966
Related Products:

sdAb anti-RFP (Cat. No. N0405)

Clone: 2B12
Host: Alpaca
Produced in: E.coli
Application:

IF (ICC)

Note: This product is not recommended for detecting proteins in Western blot, as sdAbs tend to recognize native/folded proteins mainly.

Dilution: 1:500 (corresponding to 5 nM final concentration)
Capacity: N/A
Antigen: mCherry
Targets: mRFP, mScarlet-i, mCherry
Specificity: Recognizes mScarlet-i, mScarlet-H, mScarlet-3 in its native conformation. Cross-reacts with some mRFP-derived variants like mCherry, mOrange2, DsRed1, DsRED2 and tdTomato.
Formulation:

A single sdAb clone was lyophilized from PBS pH 7.4 containing 2% BSA (US-Origin). For more details, click the Protocols button above and check Reconstitution and Storage.

kDa: -
Ext Coef: -
Shipping: Ambient temperature
Storage:

Vials containing lyophilized reagent can be stored at 2-8°C for up to 12 months. After reconstitution, store at -80°C for up to 6 months. Working aliquots can be stored at -20°C for up to 4 weeks. For more details, click the Protocols button above and check Reconstitution and Storage.

Protocols:

Relevant protocols can be found under the Protocols button above. For additional information, visit our Resources page.

References:
  1. Kashyap P, Bertelli S, Cao F, et al. An optogenetic method for the controlled release of single molecules. Nat Methods. Published online March 8, 2024. doi:10.1038/s41592-024-02204-x (IF/ICC, TIRF)
  2. Peng X, Lai KS, She P, et al. Induction of Wnt signaling antagonists and p21-activated kinase enhances cardiomyocyte proliferation during zebrafish heart regeneration [published correction appears in J Mol Cell Biol. 2022 Jan 29;13(12):921]. J Mol Cell Biol. 2021;13(1):41-58. doi:10.1093/jmcb/mjaa046 (IF/IHC; zebrafish)
  3. Mahen R, Schulte R. Cell-cell Fusion of Genome Edited Cell Lines for Perturbation of Cellular Structure and Function. J Vis Exp. 2019;(154):10.3791/60550. Published 2019 Dec 7. doi:10.3791/60550
  4. Mahen R. Stable centrosomal roots disentangle to allow interphase centriole independence. PLoS Biol. 2018;16(4):e2003998. Published 2018 Apr 12. doi:10.1371/journal.pbio.2003998 (IF)
Notice: To be used in vitro/ for research only. Non-toxic, non-hazardous, non-infectious.
Legal terms: By purchasing this product you agree to our general terms and conditions.