Multiparameter flow cytometric analysis of Granulocytes expression on Lewis Rat bone marrow cells. Lewis Rat bone marrow cells 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. 550270/550271). The cells were then stained with either FITC Mouse IgM, κ Isotype Control (Cat. No. 553474; Left Plot) or FITC Mouse Anti-Rat Granulocytes antibody (Cat. No. 554907; Right Plot) at 0.125 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of Granulocytes (or Ig Isotype control staining) versus side light-scatter signals (SSC-A) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD FACSCanto™ II Flow Cytometry System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
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Multiparameter flow cytometric analysis of Granulocytes expression on Lewis Rat bone marrow cells. Lewis Rat bone marrow cells 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. 550270/550271). The cells were then stained with either FITC Mouse IgM, κ Isotype Control (Cat. No. 553474; Left Plot) or FITC Mouse Anti-Rat Granulocytes antibody (Cat. No. 554907; Right Plot) at 0.125 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of Granulocytes (or Ig Isotype control staining) versus side light-scatter signals (SSC-A) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD FACSCanto™ II Flow Cytometry System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
Rat (QC Testing)
Mouse IgM, κ
PVG Rat Spleen Cells
Flow cytometry (Routinely Tested)
0.5 mg/ml
AB_395595
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
FITC Mouse IgM, κ Isotype Control RUO
Size
0.25 mg
Cat No.
553474
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Purified Mouse Anti-Rat CD32 RUO
Size
0.5 mg
Cat No.
550271
Cell Viability Solution RUO
Size
500 Tests
Cat No.
555815
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
554907 Rev. 10
Antibody Details
HIS48
The HIS48 antibody reacts with an antigen that is variably expressed on rat granulocytes and monocyte subsets. The HIS48 antibody also recognizes developing cells of the erythroid cell lineage. PVG rat spleen cells were used as the immunogen to generate the hybridoma that produces the HIS48 antibody.
554907 Rev. 10
Format Details
FITC
Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
FITC
Blue 488 nm
494 nm
518 nm
554907 Rev.10
Citations & References
View product citations for antibody "554907" on CiteAb
Development References (6)
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Badger DA, Sauer JM, Hoglen NC, Jolley CS, Sipes IG. The role of inflammatory cells and cytochrome P450 in the potentiation of CCl4-induced liver injury by a single dose of retinol. Toxicol Appl Pharmacol. 1996; 141(2):507-519. (Clone-specific: Immunohistochemistry). View Reference
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Barnett-Vanes A, Sharrock A, Birrell MA, Rankin S. A Single 9-Colour Flow Cytometric Method to Characterise Major Leukocyte Populations in the Rat: Validation in a Model of LPS-Induced Pulmonary Inflammation.. PLoS One. 2016; 11(1):e0142520. (Clone-specific: Flow cytometry). View Reference
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Dolen Y, Gunaydin G, Esendagli G, Guc D. Granulocytic subset of myeloid derived suppressor cells in rats with mammary carcinoma.. Cell Immunol. 2015; 295(1):29-35. (Clone-specific: Flow cytometry). View Reference
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Kiefer J, Zeller J, Bogner B, et al. An Unbiased Flow Cytometry-Based Approach to Assess Subset-Specific Circulating Monocyte Activation and Cytokine Profile in Whole Blood.. Front Immunol. 2021; 12:641224. (Clone-specific: Flow cytometry). View Reference
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Zhou X, Zhang C, Wu X, et al. Dusp6 deficiency attenuates neutrophil-mediated cardiac damage in the acute inflammatory phase of myocardial infarction.. Nat Commun. 2022; 13(1):6672. (Clone-specific: Flow cytometry). View Reference
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van Goor H, Fidler V, Weening JJ, Grond J. Determinants of focal and segmental glomerulosclerosis in the rat after renal ablation. Evidence for involvement of macrophages and lipids. Lab Invest. 1991; 64(6):754-765. (Clone-specific: Immunohistochemistry). View Reference
554907 Rev. 10
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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.