Flow cytometric analysis of CD14 expression on Rhesus macaque peripheral blood monocytes. Rhesus macaque whole blood was stained with the APC-H7 Mouse anti-Human CD14 antibody (Cat. No. 561384; solid line histogram) or with an APC-H7 Mouse IgG2a, κ Isotype Control (Cat. No. 560897; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD14 expression on Rhesus macaque peripheral blood monocytes. Rhesus macaque whole blood was stained with the APC-H7 Mouse anti-Human CD14 antibody (Cat. No. 561384; solid line histogram) or with an APC-H7 Mouse IgG2a, κ Isotype Control (Cat. No. 560897; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
LPS receptor; LPS-R; Myeloid cell-specific leucine-rich glycoprotein
Rhesus, Cynomolgus, Baboon (QC Testing)
Mouse IgG2a, κ
Human CD14 Protein
Flow cytometry (Routinely Tested)
5 µl
II M34; III M329
929
AB_10611720
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with APC-H7 under optimum conditions, and unconjugated antibody and APC-H7 were removed.
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.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- Cy is a trademark of Amersham Biosciences Limited.
- BD APC-H7 is a tandem conjugate and an analog of APC-Cy7 with the same spectral properties. It has decreased intensity but it is engineered for greater stability and less spillover in the APC channel and consequently offers better performance than APC-Cy7. It has an absorption maximum of approximately 650 nm. When excited by light from a red laser, the APC fluorochrome can transfer energy to the cyanine dye, which then emits at a longer wavelength. The resulting fluorescent emission maximum is approximately 767 nm. BD recommends that a 750-nm longpass filter be used along with a red-sensitive detector such as the Hamamatsu R3896 PMT. As with APC-Cy7 special filters are required when using APC-H7 in conjunction with APC. Note: Although our APC-H7 products demonstrate higher lot-to lot consistency than other APC tandem conjugate products, and every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-H7 conjugate.
- Although BD APC-H7 is engineered to minimize spillover to the APC channel and is more stable and less affected by light, temperature, and formaldehyde-based fixatives, compared to other APC-cyanine tandem dyes, it is still good practice to minimize as much as possible, any light, temperature and fixative exposure when working with all fluorescent conjugates.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
APC-H7 Mouse IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560897
561384 Rev. 1
Antibody Details
M5E2
The M5E2 monoclonal antibody specifically binds to CD14, a 53–55 kDa glycosylphosphatidylinositol (GPI)-anchored single chain glycoprotein expressed at high levels on monocytes. Additionally, the anti-CD14 antibody reacts with interfollicular macrophages, reticular dendritic cells, and some Langerhans cells. CD14 has been identified as a high affinity cell-surface receptor for complexes of lipopolysaccharide (LPS) and serum LPS-binding protein, LPB.
561384 Rev. 1
Format Details
APC-H7
The BD Horizon™ APC-H7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 659 nm and an acceptor dye, H7, with an emission maximum (Em Max) at 782 nm. APC-H7, driven by BD innovation, is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-H7
Red 627-640 nm
659 nm
782 nm
561384 Rev.1
Citations & References
View product citations for antibody "561384" on CiteAb
Development References (3)
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:1-1050.
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Wright SD, Ramos RA, Tobias PS, Ulevitch RJ, Mathison JC. CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science. 1990; 249(4975):1431-1433. (Biology). View Reference
561384 Rev. 1
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.