Chromeo™ 494 Goat anti-Rabbit IgG

Catalog No: 15042 Format: 1 mg $185 Buy
Catalog No: 15062 Format: 100 µg $105 Buy
Application:Immunofluorescence
Absorption:489 nm
Emission:624 nm
Molar Extinction:55,000 M-1 cm-1
Quantum Yield:~7%
Antibody Type:Polyclonal
Isotype:IgG
Host:Goat
Reactivity:Rabbit
Purification:Affinity Purified

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Contents

Contents: 1 mg (Catalog No. 15042) or 0.1 mg (Catalog No. 15062) of Chromeo™ 494 conjugated Goat anti-Rabbit IgG (H+L). The antibody concentration is 2 mg/ml in 0.01 M potassium phosphate, 0.15 M sodium chloride pH 7.4, containing 2 mM sodium azide.

Specificity: This antibody was purified by immunoaffinity chromatography. It reacts with whole molecular rabbit IgG and the light chains of other rabbit immunoglobulins. No cross-reactivity with non-immunoglobulin serum proteins was observed.

Chromeo 494 is a unique dye with special spectral properties. It is excitable with a green laser and has a large Stokes shift of 134 nm. Chromeo 494 conjugated antibodies exhibit superior luminescent properties and stability towards photobleaching, and have absorption and emission maxima of approximately 494 and 628 nm, making them ideal partners for other 488 nm excitable dyes and conjugates.

Application Notes

Applications:
Immunofluorescence: 1:100 to 1:250 dilution
Plate-based assays: 1:800 to 1:1,000 dilution

To ensure a maximum in photostability of Chromeo 494 conjugates under all experimental conditions, we recommend the use of Mowiol or TDE as mounting medium. Vectashield or MAXfluor Mounting medium might negatively influence the photostability of Chromeo 494.

Chromeo 494 Goat anti-Rabbit IgG is recommended by Leica Microsystems for Super-Resolution Fluorescence Microscopy using Leica's STimulated Emission Depletion (STED) microscopes. STED enables resolution of 50-70 nm.

 
Absorption and emission spectra of Chromeo 494 Dye.

Absorption and emission spectra of Chromeo 494 Dye.

HeLa cells stained with alpha Tubulin pAb and Chromeo 494 Goat anti-Rabbit IgG.

HeLa cells stained with alpha Tubulin pAb and Chromeo 494 Goat anti-Rabbit IgG.
HeLa cells were stained with alpha Tubulin rabbit pAb and Chromeo 494 Goat anti-Rabbit IgG.

Multi-color staining of Phospho H2AX and tubulin using Chromeo 494 and Chromeo 488 secondary antibodies in untreated and etoposide-treated HeLa cells.

Multi-color staining of Phospho H2AX and tubulin using Chromeo 494 and Chromeo 488 secondary antibodies in untreated and etoposide-treated HeLa cells.
HeLa cells were either left untreated or treated with 100 µM etoposide for 6 hours prior to fixation with methanol. The histone-variant H2AX was stained with Histone H2AX phospho Ser139 rabbit pAb (Cat. No. 39117) and Chromeo 494 Goat anti-Rabbit IgG, while tubulin was visualized using alpha Tubulin mouse mAb (Clone 5-B-1-2, Cat. No. 39527) and Chromeo 488 Goat anti-Mouse IgG (Cat. No. 15031). The nuclei were counter-stained with DAPI, then the three separate images were merged.

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.