Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 1H7 monoclonal antibody specifically binds to the α subunit of the human insulin-like growth factor-I (IGF-I) receptor that is also known as CD221. CD221 is a receptor tyrosine kinase that has high affinity for IGF-1 and low affinity for insulin and IGF-2. CD221 is synthesized as a 180 kDa precursor molecule that is processed into two subunits, a 135 kDa α subunit which is disulfide-linked to a 90 kDa β subunit. The mature IGF-IR complex is comprised of two extracellular α subunits and two transmembrane β subunits that each possesses a tyrosine kinase domain. When bound by ligand, the IGF-1 receptor autophosphorylates multiple tyrosine sites in its β chains and phosphorylates other signaling molecules that regulate cell growth, development, and neoplastic transformation. CD221 is structurally and functionally similar to the insulin receptor (aka, CD220) and is expressed on a variety of human hematopoietic and non-hematopoietic cells. The 1H7 antibody does not crossreact with the human insulin receptor and can block IGF-I- or IGF-II-stimulated cell growth.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.
Development References (3)
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Kooijman RK, Scholtens LE, Rijkers GT, Zegers BJ. Differential expression of type I insulin-like growth factor receptors in different stages of human T cells. Eur J Immunol. 1995; 25(4):931-935. (Biology). View Reference
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Li SL, Kato J, Paz IB, Kasuya J, Fujita-Yamaguchi Y. Two new monoclonal antibodies against the alpha subunit of the human insulin-like growth factor-I receptor. Biochem Biophys Res Commun. 1993; 196(1):92-98. (Clone-specific: Blocking, Inhibition, Western blot). View Reference
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Xiong L, Kasuya J, Li SL, Kato J, Fujita-Yamaguchi Y. Growth-stimulatory monoclonal antibodies against human insulin-like growth factor I receptor. Proc Natl Acad Sci U S A. 1992; 89(12):5356-5360. (Biology). View Reference
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.
