One-Step Antibody Biotinylation

Home , Biotinylation

140-002-196.04 1. μg of antibody. 10 of antibody. μg The antibody is simply added to the lyophilized biotinylation mix mix biotinylation lyophilized the to added simply is antibody The rapid the for is optimized Kit Biotinylation Antibody One-step The 1.1 1. Description This product is for research use only.use is forresearch product This

Contents 2. Capacity Components 3. Each well contains enough label for the optimal biotinylation of biotinylation optimal for the label enough contains well Each directly used in downstream applications, no post-conjugation no post-conjugation applications, downstream in used directly choice, after which the biotinylated antibody can be directly used. directly be can antibody the biotinylated which after choice, Product format [email protected] www.miltenyibiotec.com www.miltenyibiotec.com and incubated at room temperature. The labeled antibody can be can antibody labeled The temperature. at room incubated and of antibody the with incubation and resuspension simple the allows agent labeling lyophilized The of antibodies. biotinylation easy and Storage purification is necessary. is necessary. purification Phone Phone Friedrich-Ebert-Straße Friedrich-Ebert-Straße Miltenyi Biotec B.V. KG &Co. Biotec Miltenyi Protocol Description Principle of the One-step Antibody Biotinylation Kit Biotinylation Antibody One-step of the Principle Examples of immunofluorescent staining with biotinylated biotinylated with staining of immunofluorescent Examples 1.4 Applications 1.3 1.2 1.1 antibodies +49 2204 8306-0, 2204 +49

Reagent requirements Reagent information Background Kit Biotinylation Antibody One-step of the Principle

Store at –20 The labeling agent is supplied as a lyophilized lyophilized as a supplied is agent labeling The of up10 to reactions For 8labeling 1 strip of 8 reaction wells, each containing containing each wells, of 8reaction 1 strip well can be cut from the strip for individual for individual strip the from cut be can well each. use or multiple antibodies can be labeled labeled be can antibodies or multiple use simultaneously. simultaneously. the bag label. bag the for the apportioned optimally is and powder labeling of up10 to labeling lyophilized Biotinylation Mix. Biotinylation lyophilized before use. The expiration date is indicated on indicated is date expiration The use. before 68, 5142968, Fax Fax +49 2204 85197 2204 +49 Bergisch Gladbach, Germany Bergisch Gladbach, °C. Warm kit to room temperature temperature room to Warm kit °C. μg of antibody per well. Each Each well. per of antibody μg μg of antibody of antibody μg Anti-Biotin MicroBeads (# MicroBeads Anti-Biotin The One-step Antibody Biotinylation Kit has been developed for developed been has Kit Biotinylation Antibody One-step The 1.2 Applications Applications 1.3 1.4 Biotinylation Kit Antibody One-step

● ● ● ● Once biotinylated, antibodies can be directly used to tag target cells. cells. target tag to used directly be can antibodies biotinylated, Once with a FITC-, PE-, or APC-conjugated anti-biotin antibody. After After antibody. anti-biotin PE-, or APC-conjugated aFITC-, with Figure 1: Immunomagnetic labeling principle. labeling 1: Immunomagnetic Figure Subsequently, target cells can either be magnetically labeled using using labeled magnetically be either can cells target Subsequently, separation (with MACS® Technology) as well as fluorescent cell cell fluorescent as well Technology) as MACS® (with separation analysis. according to the protocol in the Anti-Biotin MicroBeads data sheet. data MicroBeads Anti-Biotin the in protocol the to according page 1/2 the biotinylation of monoclonal antibodies for use in magnetic cell cell magnetic in for use antibodies of monoclonal biotinylation the labeling with Anti-Biotin MicroBeads, target cells can be enriched enriched be can cells target MicroBeads, Anti-Biotin with labeling

Background information Background Biotinylation of monoclonal antibodies. antibodies. of monoclonal Biotinylation (Optional) Anti-Biotin-FITC (# (# 130-090-485). MicroBeads Anti-Biotin (Optional) be purified must be biotinylated to antibody The Antibody: Reagent requirements Reagent (# μg of antibody can be labeled per well. per labeled be can of 10 of antibody μg Biotin-VioBlue® (# molecules prior to biotinylation. The antibody should be should antibody The biotinylation. to prior molecules NH₂-containing other and components, serum azide, from 094-974) for use as a secondary antibody reagent. antibody asecondary 094-974) as for use prepared at a concentration of 100 at aconcentration prepared 130-090-756), Anti-Biotin-APC (# Anti-Biotin-APC 130-090-756), primary antibody primary Biotinylated MicroBead Anti-Biotin 130-094-669), or Anti-Biotin-PerCP (# or Anti-Biotin-PerCP 130-094-669), 130-090-485) or fluorescently stained stained or fluorescently 130-090-485)

130-090-857), Anti-Biotin-PE μg/mL in PBS. A maximum Amaximum PBS. in μg/mL Order no. 130-093-385 no. Order 130-090-856), Anti- 130-090-856), 130- Order no. 130-093-385

2. General protocol for immunofluorescent staining Refer to www.miltenyibiotec.com for all data sheets and protocols. Miltenyi Biotec provides technical support worldwide. Visit ▲ The following protocol is for the biotinylation of 100 μL of a www.miltenyibiotec.com for local Miltenyi Biotec Technical single antibody adjusted to a concentration of 100 μg/mL in PBS. Support contact information. As desribed above, the antibody must be free of azide, serum components, and other NH₂-containing molecules. 1. Cut off and thaw the required number of wells from the strip. Wells must be warmed to room temperature before use. Legal notices

Limited product warranty 2. Add 100 μL of the antibody to be biotinylated to the well. Miltenyi Biotec B.V. & Co. KG and/or its affiliate(s) warrant this product to be free Resuspend the lyophilized powder fully by mixing thoroughly. from material defects in workmanship and materials and to conform substantially with Miltenyi Biotec’s published specifications for the product at the time of order, 3. Incubate the mixture at a controlled room temperature under normal use and conditions in accordance with its applicable documentation, (18–25 °C) for 24 h. for a period beginning on the date of delivery of the product by Miltenyi Biotec or its authorized distributor and ending on the expiration date of the product’s applicable 4. After incubation, the antibody is ready for direct use in cell shelf life stated on the product label, packaging or documentation (as applicable) or, labeling or other downstream applications. in the absence thereof, ONE (1) YEAR from date of delivery (“Product Warranty”). ▲ Note: Optimal staining concentrations must be determined Miltenyi Biotec’s Product Warranty is provided subject to the warranty terms as set forth in Miltenyi Biotec’s General Terms and Conditions for the Sale of Products and ▲ Note: Store conjugated antibodies at 2–8 °C. For long-term storage, addition Services available on Miltenyi Biotec’s website at www.miltenyibiotec.com, as in effect of preservative is recommended. at the time of order (“Product Warranty”). Additional terms may apply. BY USE OF ▲ Note: Avoid use of streptavidin or Streptavidin MicroBeads as a second step THIS PRODUCT, THE CUSTOMER AGREES TO BE BOUND BY THESE TERMS. reagent as residual free biotin in the labeling mixture might cause increased THE CUSTOMER IS SOLELY RESPONSIBLE FOR DETERMINING IF A PRODUCT background staining. Free biotin is not detected by Anti-Biotin MicroBeads and IS SUITABLE FOR CUSTOMER’S PARTICULAR PURPOSE AND APPLICATION -antibodies METHODS.

3. Examples of immunofluorescent staining with Technical information The technical information, data, protocols, and other statements provided by Miltenyi biotinylated antibodies Biotec in this document are based on information, tests, or experience which Miltenyi Biotec believes to be reliable, but the accuracy or completeness of such information With the One-step Antibody Biotinylation Kit, 10 μg of unconjugated is not guaranteed. Such technical information and data are intended for persons with CD15 and CD4 antibodies were biotinylated as described. Human knowledge and technical skills sufficient to assess and apply their own informed peripheral blood mononuclear cells (PBMCs) were then stained with judgment to the information. Miltenyi Biotec shall not be liable for any technical or the biotinylated antibodies as well as Anti-Biotin-PE (# 130-090- editorial errors or omissions contained herein. 756) (A, B). Cells labeled with biotinylated CD4 (VIT4) were also All information and specifications are subject to change without prior notice. Please contact Miltenyi Biotec Technical Support or visit www.miltenyibiotec.com for the stained with CD14-FITC (B). Cells were analyzed by flow cytometry. most up-to-date information on Miltenyi Biotec products. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. Licenses This product and/or its use may be covered by one or more pending or issued patents and/or may have certain limitations. Certain uses may be excluded by separate terms and conditions. Please contact your local Miltenyi Biotec representative or visit A) Miltenyi Biotec’s website at www.miltenyibiotec.com for more information. The purchase of this product conveys to the customer the non-transferable right to use the purchased amount of the product in research conducted by the customer (whether the customer is an academic or for-profit entity). This product may not be further sold. Additional terms and conditions (including the terms of a Limited Use Label License) may apply. CUSTOMER’S USE OF THIS PRODUCT MAY REQUIRE ADDITIONAL LICENSES DEPENDING ON THE SPECIFIC APPLICATION. THE CUSTOMER IS SOLELY RESPONSIBLE FOR DETERMINING FOR ITSELF WHETHER IT HAS ALL APPROPRIATE LICENSES IN PLACE. Miltenyi Biotec provides no warranty that

CD15-Biotin/Anti-Biotin-PE customer’s use of this product does not and will not infringe intellectual property Forward scatter rights owned by a third party. BY USE OF THIS PRODUCT, THE CUSTOMER AGREES TO BE BOUND BY THESE TERMS.

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CD14-FITC

Unless otherwise specifically indicated, all Miltenyi Biotec products and services page 2/2 are for research use only and not for diagnostic or therapeutic use. 140-002-196.04