- FC, IF
- Description of Target:
Mix-n-Stain™ antibody labeling kits contain everything you need to rapidly label an antibody with Aviva's next-generation CF™ dyes or biotin. The labeling procedure comprises simple mixing of your antibody with the reaction buffer and optimally formulated dye provided, followed by a brief incubation. The Mix-n-Stain dye is no longer reactive at the end of the labeling, so the conjugate is ready for staining without further purification. After labeling, the dye is covalently linked to the antibody with a degree of labeling of approximately 4-6 dye molecules per antibody molecule. Mix-n-Stain labeling is covalent, so Mix-n-Stain-labeled antibodies can be used for multicolor fluorescence staining without transfer of dyes between antibodies.
Mix-n-Stain labeling can tolerate sodium azide, and low levels of glycerol, Tris, and glycine. A microcentrifuge ultrafiltration vial is provided in the kit to rapidly remove incompatible small molecule antibody stabilizers before labeling.
The standard Mix-n-Stain labeling protocol can be performed in the presence of up to four-fold excess of BSA or gelatin to IgG (by ug amount). Simply choose the kit size that corresponds to the amount of IgG you wish to label. A modified protocol is provided for labeling IgG in the presence of excess stabilizer protein or ascites fluid.
In this case, choose the kit size that corresponds to the total amount of protein (IgG + stabilizer, or total protein amount in ascites fluid) in the antibody sample you wish to label. The modified protocol also can be used to label amounts of IgG that fall below the lower range of the kit by adding stabilizer protein to the IgG to bring the total protein amount within the kit range. The modified protocol is not recommended for labeling antibodies in crude antiserum or hybridoma cell culture supernatant due to the low concentration of antibody relative to total protein in these formats.
When performing direct immunofluorescence with a fluorescently-labeled antibody, you may need to use a higher concentration of antibody to achieve similar staining intensity compared to indirect immunofluorescence detection using unlabeled primary plus labeled secondary antibody. In our internal testing, indirect immunofluorescence staining results in about 3-fold signal amplification compared to direct immunofluorescence staining.
Labeled secondary antibodies will still bind to primary antibodies labeled using Mix-n-Stain kits, therefore if multiple primary antibodies from the same species are to be used for multicolor immunofluorescence staining, a secondary antibody cannot be used to distinguish an unlabeled primary antibody from a Mix-n-Stain labeled primary antibody from the same species. Mix-n-Stain labeled antibodies can be used as a tertiary staining antibody after standard immunofluorescence staining with primary and secondary antibodies. Aviva also offers biotin Mix-n-Stain labeling kits, for secondary detection using CF™ dye-labeled streptavidin or CF™ dye-labeled monoclonal mouse anti-biotin.
- Printable datasheet/reference manual for Mix-n-Stain CF640R Antibody Labeling Kit,1x(20-50ug) labeling (OKBE00036) OKBE00036
- Reconstitution and Storage:
Stability: Stable for at least 3 months from date of receipt when stored as recommended.
- Additional Information:
- Technical Summary
Abs/Em Maxima: 642/662 nm
Extinction coefficient: 105,000
Molecular weight: ~ 832
Excitation laser line: 633 nm, 635 nm or 640 nm
Direct replacement for: Alexa Fluor® 647, ATTO™ 647N, Cy™5, DyLight® 649
-Has the best photostability among dyes with Cy®5-like spectra
-Yields highly fluorescent protein conjugates
-Very water-soluble and pH-insensitive
|Component|| 20-50 ug kit|
|Dye vial ||(1) vial Component A|
|Mix-n-Stain reaction|| (1) vial of 15 uL buffer, 10X Part # 99951-1|
|Mix-n-Stain antibody ||(1) vial of 150 uL storage buffer Part # 99952-1|
|Ultrafiltration vial (MWCO=10K) ||1 each Part # 99956|
- Please visit www.avivasysbio.com to view our full selection of products featuring bright and photostable CF™ dyes, including secondary antibodies, streptavidin, anti-biotin, and anti-tag antibodies. Aviva also offers a variety of apoptosis and cell viability assays for flow cytometry analysis, including mitochondrial membrane potential dyes, fluorescent Annexin V conjugates, and NucView™488 Caspase-3 Substrate for live cells.
CF dye, Mix-n-Stain, and modified Mix-n-Stain labeling technologies are covered by pending US and international patents. TWEEN is a registered trademark of Uniqema Americas LLC.
Aviva products are high-quality reagents and materials intended for research purposes only. Some products are potentially hazardous chemicals - please read the Material Safety Data Sheet for additional information regarding handling potentially hazardous chemicals. Several of Aviva products and product applications are covered by U.S. and international patents and pending patents. Our products are not available for resale or other commercial uses without a specific agreement from Aviva Systems Biology Corporation. We welcome inquiries about licensing the use of our dyes, trademarks or technologies. Please submit inquiries by e-mail to email@example.com
- Mix-n-StainTM Antibody Compatibility and Labeling Protocol Selection Guide
|Glycerol||< 10%: proceed to standard protocol (Section B)|
> 10%: perform ultrafiltration (Section A)
|Tris||< 20 mM: proceed to standard protocol (Section B)|
> 20 mM: perform ultrafiltration (Section A)
|Glycine||Perform ultrafiltration (Section A)|
|BSA or gelatin||< 4X IgG by ug amount: use standard protocol (Section B)|
> 4X IgG by ug amount: use modified protocol (Section C)
|Ascites fluid||Use modified protocol (Section C)|
|Serum||Not compatible; purify IgG|
|Not compatible: purify IgG|
- Tips Information: