Invitrogen

CD14 Monoclonal Antibody (61D3), Super Bright™ 600, eBioscience™

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CD14 Antibody in Flow Cytometry (Flow) — Staining of normal human peripheral blood cells with Mouse IgG1 K Isotype Control Super Bright 600 (Product # 63-4714-82) (blue histogram) or Anti-Human CD14 Super Bright 600 (purple histogram). Cells in the monocyte gate were used for analysis.

CD14 Antibody in Immunocytochemistry (ICC/IF) — Figure 1 PMA-induced TM expression mediates morphological changes and differentiation marker expression in THP-1 cells. ( A ) THP-1 cells were treated with 150 nM PMA for 24-72 hours. The total cell lysates were harvested, and the expression of TM was analyzed using western blot analysis. beta-actin was used as a loading control. Five independent experiments have been performed (n = 5) and representative images have been showed. The amount of proteins expression was quantified using densitometry and presented as bar graph. The data are presented as the mean +- SD (n = 5), and * p < 0.05 was considered significant. ( B ) THP-1 cells were transfected with TM siRNA or HA-TM FL plasmid for 24 h followed by PMA stimulation for 72 hours. The morphology of the cells was observed using light microscopy. The adherent differentiated macrophage-like cells are indicated by a white arrowhead. Five independent experiments have been performed (n = 5). The quantification is shown in the right graph. ( C ) The expression of the macrophage cell surface markers CD14 (red) and CD68 (green) was analyzed using immunofluorescence and microscopy. Hoechst staining was used to label the nuclei. The scale bar indicates 100 mum. Five independent experiments have been performed (n = 5), and showed representative images. ( D ) The expression of CD14 and CD68 was analyzed using flow cytometry. Data are expressed as a % of the control, are presented as the mean +- SD and represent the results of three indep

CD14 Antibody in Immunohistochemistry (IHC) — Figure 2 CD14 and TLR-4 staining in gut epithelial cells. Balb/c mice were not supplemented (control) or supplemented daily by oral gavage with OxC-beta (10 mg/kg). After 4 weeks, intestinal tissues were harvested and CD14 and TLR-4 expression was determined by immunocytochemistry. Increased CD14 (A) and TLR-4 (C) expression is readily apparent in epithelial cells in the OxC-beta-supplemented animals compared to the controls receiving vehicle alone (B and D, respectively). Arrows indicate the location of enterocytes within the cross section of microvilli. Magnification 40x.

产品信息

63-0149-42

应用	建议稀释比	已发表文章
免疫组化 (IHC)	-	查看1篇已发表文章 1篇已发表文章
免疫细胞化学 (ICC/IF)	-	查看1篇已发表文章 1篇已发表文章
流式细胞分析 (Flow)	5 µL (0.5 µg)/test	查看6篇已发表文章 6篇已发表文章

产品规格
种属反应	Human
已发表种属	Not Applicable
宿主/亚型	Mouse / IgG1, kappa
推荐的同型对照	Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), Super Bright™ 600, eBioscience™
分类	Monoclonal
类型	Antibody
克隆号	61D3
偶联物	Super Bright™ 600 Alexa Fluor 488 Alexa Fluor 532 Alexa Fluor 660 Alexa Fluor 700 APC APC-eFluor 780 Biotin Brilliant UV 395 Brilliant UV 496 Brilliant UV 615 Brilliant UV 737 Brilliant UV 805 Brilliant Violet 421 eFluor 450 eFluor 506 FITC Functional Grade NovaFluor Blue 510 NovaFluor Blue 555 NovaFluor Blue 585 NovaFluor Blue 610-30S NovaFluor Blue 660-40S NovaFluor Yellow 570 NovaFluor Yellow 660
激发/发射光谱	查看光谱
形式	Liquid
浓度	5 µL/Test
纯化类型	Affinity chromatography
保存液	PBS, pH 7.2, with BSA
内含物	0.09% sodium azide
保存条件	4° C, store in dark, DO NOT FREEZE!
RRID	AB_2662456

产品详细信息

Description: The 61D3 monoclonal antibody reacts with human CD14, a 53-55 kDa GPI-linked glycoprotein. CD14 is expressed on monocytes, interfollicular macrophages and some dendritic cells. Complexes of LPS and LBP (LPS-Binding Protein) bind with high affinity to monocytes through the surface CD14.

Applications Reported: This 61D3 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 61D3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10⁵ to 10⁸ cells/test.

Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 2-8°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorochrome performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.

靶标信息

CD14 is a 55 kDa GPI-anchored glycoprotein that is constitutively expressed on the surface of mature monocytes, macrophages, and neutrophils. CD14 also serves as a multifunctional lipopolysaccharide receptor, and is released to the serum both as a secreted and enzymatically cleaved GPI-anchored form. CD14 binds lipopolysaccharide molecule in a reaction catalyzed by lipopolysaccharide-binding protein (LBP), an acute phase serum protein. The soluble sCD14 can discriminate slight structural differences between lipopolysaccharides and is important for neutralization of serum allochthonous lipopolysaccharides by reconstituted lipoprotein particles. Further, CD14 has been shown to bind apoptotic cells, and can affect allergic, inflammatory and infectious processes. Alternative splicing results in multiple transcript variants encoding the same CD14 isoform. Diseases associated with CD14 dysfunction include mycobacterium chelonae infection and Croup.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

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