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Phospho Btk (Y551) Flex Set

BD™ Cytometric Bead Array (CBA) Phospho Btk (Y551) Flex Set

(RUO)
Phospho Btk (Y551) Flex Set
Figure 1. Example BD CBA Phospho Btk (Y551) Flex Set standard curve. Data acquired on a BD FACSArray bioanalyzer and analyzed using FCAP Array Software.
Phospho Btk (Y551) Flex Set
Reproducibility: The inter-assay and intra-assay reproducibility were determined for the BD CBA Phospho Btk (Y551) Flex Set by evaluating ten replicates of three different sample levels (intra-assay) and two replicates of three different sample levels from four separate experiments (inter-assay).
Phospho Btk (Y551) Flex Set
Linearity: An activated cell lysate was serially diluted to determine the linearity of the assay.
Figure 1. Example BD CBA Phospho Btk (Y551) Flex Set standard curve. Data acquired on a BD FACSArray bioanalyzer and analyzed using FCAP Array Software.
Reproducibility: The inter-assay and intra-assay reproducibility were determined for the BD CBA Phospho Btk (Y551) Flex Set by evaluating ten replicates of three different sample levels (intra-assay) and two replicates of three different sample levels from four separate experiments (inter-assay).
Linearity: An activated cell lysate was serially diluted to determine the linearity of the assay.
Product Details
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BD™ Cytometric Bead Array (CBA)
D5
3.9-1,000 Units/mL
Human (QC Testing)
RUO
AB_2869269


Description

The BD™ CBA Phospho Btk (Y551) Flex Set is a bead-based immunoassay capable of measuring human Bruton's tyrosine kinase (Btk), a non-receptor tyrosine kinase, which has been tyrosine-phosphorylated at Tyr-551 in denatured cell lysate samples. Human reactivity was determined by testing cell lysates with the BD CBA Phospho Btk (Y551) Flex Set. There are several publications evaluating the biology and function of Btk. For more information on bead-based immunoassays, refer to the product insert for the BD CBA Cell Signaling Master Buffer Kit (Cat. No. 560005 or 560006).

Preparation And Storage

This BD™ CBA Flex Set contains one vial of each component listed above. All components of this flex set have been formulated to a 50x concentration to ensure product performance when multiplexed. Store at 4°C. Protect Capture Beads and the PE Detection Reagent from prolonged exposure to light.

The Phospho Btk (Y551) Standard provided in this Flex Set is lyophilized and the standard sphere should be transferred to a 1.5 mL microfuge tube for reconstitution. Reconstitute the standard with 0.1 mL Assay Diluent from the BD CBA Cell Signaling Master Buffer Kit (Cat. No. 560005/560006), warm to 37 °C and vortex prior to use. After reconstitution, the standard concentration is 50,000 Units/mL and is stable for 3 months when stored at 4 °C. When using reconstituted standard, warm to 37 °C and vortex to mix thoroughly before use.

Recommended Assay Procedures

The BD CBA Phospho Btk (Y551) Flex Set must be used in conjunction with a BD CBA Cell Signaling Master Buffer Kit (Cat. No. 560005, 100 tests, or 560006, 500 tests) and a flow cytometer. Detailed instructions on the use of this product can be found in the manual for the BD CBA Cell Signaling Master Buffer Kit. When following the directions in the Master Buffer Kit, the top standard point for the BD CBA Phospho Btk (Y551) Flex Set will be 1,000 Units/mL. An example standard curve is shown in Figure 1.

The BD CBA Phospho Btk (Y551) Flex Set cannot be used in the same assay well with the following BD CBA Cell Signaling Flex Set reagents:

Flex Set                                                                Bead Position                Catalog Number

BD CBA Phospho Akt2 (T309) Flex Set                        A5                        560146

BD CBA Phospho JNK1/2 (T202/Y204) Flex Set                B5                        560013

BD CBA Phospho Pyk2 (Y402) Flex Set                        D9                        560148

The BD CBA Phospho Btk (Y551) Flex Set should not be used in the same assay well with any non-BD CBA Cell Signaling Flex Set reagents (such as BD CBA Human or Mouse Soluble Protein Flex Sets) nor with any BD CBA Total Protein Cell Signaling Flex Set reagents. For an updated assay compatibility chart for the BD CBA Cell Signaling Flex Sets, please refer to the BD CBA Flex Set System homepage at http://www.bdbiosciences.com/cbasetup.

Performance

Limit of Detection: The theoretical limit of detection is 1.88 Units/mL and was determined by evaluating the estimated result of the average MFI of the negative control (0 Units/mL, n=30) + 2 standard deviations.

Product Notices

  1. ProClin is a trademark of Rohm and Haas Company.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Warning: CBA lyophilized standard contains 32.07% sodium dodecyl sulfate (w/w) and 0.01% (w/w) of a CMIT/MIT mixture (3:1), which is a mixture of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC No 220-239-6] (3:1).Hazard statement: May be harmful if swallowed. Causes skin irritation. Causes serious eye irritation. May cause an allergic skin reaction.Precautionary statements: Wear protective gloves/eye protection. Wear protective clothing. Avoid breathing mist/vapours/spray. If skin irritation or rash occurs: Get medical advice/attention. IF ON SKIN: Wash with plenty of water. Dispose of contents/container in accordance with local/regional/national/international regulations.
560004 Rev. 4
Components
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Description Quantity/Size Part Number EntrezGene ID
Phospho Btk (Y551) Capture Bead D5 100 Tests (1 ea) 51-9005166 N/A
Phospho Btk (Y551) PE* Detection Reagent 100 Tests (1 ea) 51-9005167 N/A
Phospho Btk (Y551) Standard 100 Tests (1 ea) 51-9005169 N/A
560004 Rev. 4
Citations & References
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Development References (2)

  1. Mahajan S, Fargnoli J, Burkhardt AL, Kut SA, Saouaf SJ, Bolen JB. Src family protein tyrosine kinases induce autoactivation of Bruton's tyrosine kinase. Mol Cell Biol. 1995; 15:5304-5311. (Biology). View Reference
  2. Rawlings DJ, Scharenberg AM, Park H, et al. Activation of BTK by a phosphorylation mechanism initiated by SRC family kinases. Science. 1996; 271:822-825. (Biology). View Reference
560004 Rev. 4

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.