BD Pharmingen™ Alexa Fluor™ 647 Rat Anti-Mouse CD14
Clone Sa14-2 (also known as SA14-2) (RUO)
Multiparameter flow cytometric analysis of CD14 expression on viable Mouse thioglycolate-elicited peritoneal exudate cell populations. C57BL/6 Mouse peritoneal exudate cells were isolated 3 days post-stimulation by intraperitoneal injection of thioglycolate solution (Thio-PEC) and were preincubated with BD Pharmingen™ Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141].The cells were then stained with either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Plot) or Alexa Fluor™ 647 Rat Anti-Mouse CD14 antibody (Cat. No. 570368/570438; Right Plot) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of CD14 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) Thio-PECs. 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 CD14 expression on viable J774A.1 cells. Cells from the J774A (Mouse macrophage, ATCC® TIB-67™) cell line were stained with either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; dashed line histogram) or Alexa Fluor™ 647 Rat Anti-Mouse CD14 antibody (Cat. No. 570368/570438; solid line histogram) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing CD14 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD14 expression on viable Mouse thioglycolate-elicited peritoneal exudate cell populations. C57BL/6 Mouse peritoneal exudate cells were isolated 3 days post-stimulation by intraperitoneal injection of thioglycolate solution (Thio-PEC) and were preincubated with BD Pharmingen™ Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141].The cells were then stained with either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Plot) or Alexa Fluor™ 647 Rat Anti-Mouse CD14 antibody (Cat. No. 570368/570438; Right Plot) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of CD14 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) Thio-PECs. 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 CD14 expression on viable J774A.1 cells. Cells from the J774A (Mouse macrophage, ATCC® TIB-67™) cell line were stained with either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; dashed line histogram) or Alexa Fluor™ 647 Rat Anti-Mouse CD14 antibody (Cat. No. 570368/570438; solid line histogram) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing CD14 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
Cd14; LPS Receptor
Mouse (QC Testing)
Rat IgG2a, κ
Mouse CD14-transfected Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
12475
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 unreacted dye was 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- 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
Alexa Fluor® 647 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557690
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
DAPI Solution RUO
Size
1 mg
Cat No.
564907
570438 Rev. 1
Antibody Details
Sa14-2
The SA14-2 monoclonal antibody specifically recognizes mouse CD14. CD14 is an ~55-kDa glycosyl phosphatidyl inositol (GPI)-linked cell-surface glycoprotein that is encoded by Cd14. CD14 is expressed on macrophages, granulocytes, dendritic cells, hepatocytes, and Kupffer cells. CD14 is also expressed in a soluble form (sCD14). CD14 is a leucine-rich repeat-containing pattern recognition receptor which serves as coreceptor for bacterial lipopolysaccharide (LPS) from gram-negative bacteria. CD14 forms a complex with lipopolysaccharide (LPS), an endotoxin from gram-negative bacteria, with LPS-binding protein (LBP, a plasma protein). The LPS-LPB-CD14 complex can then associate with TLR4-MD-2 which transduces intracellular signals. In this way, CD14 can play roles in innate cellular anti-bacterial immune and inflammatory responses, such as through the stimulation of cellular IL-1 and TNF production, and in extreme cases, the development of endotoxic shock.
570438 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
570438 Rev.1
Citations & References
View product citations for antibody "570438" on CiteAb
Development References (3)
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Akashi S, Saitoh S, Wakabayashi Y, et al. Lipopolysaccharide interaction with cell surface Toll-like receptor 4-MD-2: higher affinity than that with MD-2 or CD14. J Exp Med. 2003; 198(7):1035-1042. (Biology). View Reference
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Ghosh M, Subramani J, Rahman MM, Shapiro LH. CD13 restricts TLR4 endocytic signal transduction in inflammation.. J Immunol. 2015; 194(9):4466-76. (Clone-specific: Flow cytometry). View Reference
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Kiyokawa T, Akashi-Takamura S, Shibata T, et al. A single base mutation in the PRAT4A gene reveals differential interaction of PRAT4A with Toll-like receptors.. Int Immunol. 2008; 20(11):1407-15. (Immunogen: Flow cytometry). View Reference
570438 Rev. 1
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.