PE Rat Anti-Mouse CD14

BD Pharmingen™ PE Rat Anti-Mouse CD14

Name: PE Rat Anti-Mouse CD14
Brand: PE Rat Anti-Mouse CD14
SKU: 569968

Clone Sa14-2 (also known as SA14-2) (RUO)

Flow cytometric analysis of CD14 expression on J774A.1 cells. Cells from the J774A (Mouse macrophage, ATCC® TIB-67™) cell line were stained with either BD Pharmingen™ PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; dashed line histogram) or BD Pharmingen™ PE Rat Anti-Mouse CD14 antibody (Cat. No. 569968/569969; solid line histogram) 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 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 (&-AAD-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ 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 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. 553141/553142].The cells were then stained with either BD Pharmingen™ PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or BD Pharmingen™ PE Rat Anti-Mouse CD14 antibody (Cat. No. 555815/555816; Right Plot) at 0.125 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) 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.

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Product Details

Brand: BD Pharmingen™

Alternative Name: Cd14; LPS Receptor

Reactivity: Mouse (QC Testing)

Isotype: Rat IgG2a, κ

Immunogen: Mouse CD14-transfected Cells

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

Entrez Gene ID: 12475

RRID: AB_3685441

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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.
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.
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

PE Rat IgG2a, κ Isotype Control RUO

Size 0.1 mg Cat No. 553930

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

569968 Rev. 2

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.

569968 Rev. 2

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

569968 Rev.2

Citations & References

View product citations for antibody "569968" on CiteAb

Development References (3)

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
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
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

569968 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.