BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-Rat CD38
Clone 14.27 (also known as CD38.14.27) (RUO)
Flow cytometric analysis of CD38 expression on Rat splenic leukocytes. Lewis Rat splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) [Cat. No. 550271]. The cells were then stained with FITC Mouse Anti-Rat CD3 antibody (Cat No. 557354) and with either Alexa Fluor™ 647 Mouse IgG2b, κ Isotype Control (Cat. No. 565378; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Rat CD38 antibody (Cat. No. 569804/569878; Right Plot) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD38 (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of CD38 expression on Rat splenic leukocytes. Lewis Rat splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) [Cat. No. 550271]. The cells were then stained with FITC Mouse Anti-Rat CD3 antibody (Cat No. 557354) and with either Alexa Fluor™ 647 Mouse IgG2b, κ Isotype Control (Cat. No. 565378; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Rat CD38 antibody (Cat. No. 569804/569878; Right Plot) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD38 (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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 BD® 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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
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- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- An isotype control should be used at the same concentration as the antibody of interest.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
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The 14.27 monoclonal antibody specifically recognizes CD38, which is also known as ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 or ADP-ribosyl cyclase 1 (ADPRC 1). CD38 a 45 kDa type II transmembrane glycoprotein that is encoded by Cd38 (CD38 molecule). CD38 is variably expressed on early hematopoietic precursors and leucocytes including immature and mature B cells as well as thymocytes, T cells, NK cells, monocytes, and macrophages. It is also expressed on non-hematopoietic cells such as epithelial cells, astrocytes, hepatic stellate cells (HSCs), and Kupffer cells. CD38 catalyzes the synthesis and hydrolysis of cyclic adenosine diphosphate ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP) which can function as secondary messengers involved in calcium mobilization. The 14.27 antibody can reportedly stimulate increased cytosolic Ca2+ levels, IL-6 production and increased expression of the adhesion molecules, VCAM-1, N-CAM, and ICAM-1 by HSCs.
Development References (3)
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Bath NM, Ding X, Wilson NA, et al. Desensitization and treatment with APRIL/BLyS blockade in rodent kidney transplant model.. PLoS One. 2019; 14(2):e0211865. (Clone-specific: Flow cytometry). View Reference
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March S, Graupera M, Rosa Sarrias M, et al. Identification and functional characterization of the hepatic stellate cell CD38 cell surface molecule.. Am J Pathol. 2007; 170(1):176-87. (Immunogen: Calcium Flux, Flow cytometry, Fluorescence microscopy, Functional assay, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western blot). View Reference
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Pearse DD, Bastidas J, Izabel SS, Ghosh M. Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury.. Int J Mol Sci. 2018; 19(9):E2550. (Clone-specific: Flow cytometry). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.