Dear Customer, If you experience any text readability or display issues on our website, please update your web browser to the latest version
See More
BD Pharmingen™ PE Mouse Anti-Human MIP-1β
克隆 D21-1351 (RUO)
Expression of MIP-1β by stimulated human monocytes. Human PBMC were stimulated with human IFN-γ (20 ng/ml final concentration; Cat. No. 554616/554617) for one hour followed by overnight stimulation with LPS (1 µg/ml final concentration; Sigma, Cat. No. L-8272) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, fixed, permeabilized, and stained with 0.03 µg of PE Mouse Anti-Human MIP-1β (Cat. 550078/561120; Left Panel) following Pharmingen's staining protocol. The data reflects gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, binding by the PE Mouse Anti-Human MIP-1β antibody was blocked by preincubation with recombinant human MIP-1β (0.25 µg; Middle panel) and by preincubation of the fixed/permeabilized cells with excess unlabeled D21- 1351 antibody (5 µg; custom order; Right panel) prior to staining with PE Mouse Anti-Human MIP-1β. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified using the unlabeled antibody blocking control.
Expression of MIP-1β by stimulated human monocytes. Human PBMC were stimulated with human IFN-γ (20 ng/ml final concentration; Cat. No. 554616/554617) for one hour followed by overnight stimulation with LPS (1 µg/ml final concentration; Sigma, Cat. No. L-8272) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, fixed, permeabilized, and stained with 0.03 µg of PE Mouse Anti-Human MIP-1β (Cat. 550078/561120; Left Panel) following Pharmingen's staining protocol. The data reflects gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, binding by the PE Mouse Anti-Human MIP-1β antibody was blocked by preincubation with recombinant human MIP-1β (0.25 µg; Middle panel) and by preincubation of the fixed/permeabilized cells with excess unlabeled D21- 1351 antibody (5 µg; custom order; Right panel) prior to staining with PE Mouse Anti-Human MIP-1β. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified using the unlabeled antibody blocking control.
Expression of MIP-1β by stimulated human monocytes. Human PBMC were stimulated with human IFN-γ (20 ng/ml final concentration; Cat. No. 554616/554617) for one hour followed by overnight stimulation with LPS (1 µg/ml final concentration; Sigma, Cat. No. L-8272) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, fixed, permeabilized, and stained with 0.03 µg of PE Mouse Anti-Human MIP-1β (Cat. 550078/561120; Left Panel) following Pharmingen's staining protocol. The data reflects gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, binding by the PE Mouse Anti-Human MIP-1β antibody was blocked by preincubation with recombinant human MIP-1β (0.25 µg; Middle panel) and by preincubation of the fixed/permeabilized cells with excess unlabeled D21- 1351 antibody (5 µg; custom order; Right panel) prior to staining with PE Mouse Anti-Human MIP-1β. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified using the unlabeled antibody blocking control.
Expression of MIP-1β by stimulated human monocytes. Human PBMC were stimulated with human IFN-γ (20 ng/ml final concentration; Cat. No. 554616/554617) for one hour followed by overnight stimulation with LPS (1 µg/ml final concentration; Sigma, Cat. No. L-8272) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, fixed, permeabilized, and stained with 0.03 µg of PE Mouse Anti-Human MIP-1β (Cat. 550078/561120; Left Panel) following Pharmingen's staining protocol. The data reflects gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, binding by the PE Mouse Anti-Human MIP-1β antibody was blocked by preincubation with recombinant human MIP-1β (0.25 µg; Middle panel) and by preincubation of the fixed/permeabilized cells with excess unlabeled D21- 1351 antibody (5 µg; custom order; Right panel) prior to staining with PE Mouse Anti-Human MIP-1β. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified using the unlabeled antibody blocking control.
商品详情
BD Pharmingen™
Macrophage inflammatory protein 1-beta; CCL4; C-C motif chemokine 4; LAG-1
Human (QC Testing)
Mouse IgG1, κ
Recombinant Human MIP-1β
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_393549
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
准备和存储
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
推荐的实验流程
IF/Flow: For immunofluorescent staining and flow cytometric analysis, the D21-1351 antibody has been found useful to identify and enumerate MIP-1β producing cells within mixed cell populations. PE Mouse Anti-Human MIP-1β (Cat. No. 550078/561120) is especially suitable for these studies.
A suitable mouse IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponinpermeabilized human cells is PE Mouse IgG1, κ Isotype Control (Cat. No. 554680); use at comparable concentrations to antibody of interest (e.g., . 0.5 µg mAb/1 million cells). A useful control for demonstrating specificity of staining is to pre-block the fixed/permeabilized cells with unlabeled D21-1351 antibody (custom order), prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.
商品通知
- 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 www.bdbiosciences.com/us/s/resources for technical protocols.
561120 Rev. 2
抗体详情
D21-1351
The D21-1351 monoclonal antibody specifically binds to the human CC chemokine, MIP-1β (macrophage inflammatory protein-1β). Human MIP-1β shares approximately 75% homology with mouse MIP-1β at the amino acid level. Expression of MIP-1β in human peripheral blood cells is induced by proinflammatory and mitogenic stimuli. MIP-1β is a chemoattractant for monocytes and lymphocytes. Human MIP-1β binds to receptors, CCR5 and CCR8. The human MIP-1β gene has been mapped to chromosome 17q11. The immunogen used to generate D21-1351 hybridoma was recombinant human MIP-1β.
561120 Rev. 2
格式详情
PE
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.
PE
496 nm, 566 nm
576 nm
561120 Rev.2
报价单和参考
View product citations for antibody "561120" on CiteAb
研发参考 (9)
-
Bernardini G, Hedrick J, Sozzani S. Identification of the CC chemokines TARC and macrophage inflammatory protein-1 beta as novel functional ligands for the CCR8 receptor. J Immunol. 1998; 28(2):582-588. (Biology). 查看参考
-
Combadiere C, Ahuja SK, Tiffany HL, Murphy PM. Cloning and functional expression of CC CKR5, a human monocyte CC chemokine receptor selective for MIP-1(alpha), MIP-1(beta), and RANTES. J Leukoc Biol. 1996; 60(1):147-152. (Biology). 查看参考
-
Lipes MA, Napolitano M, Jeang KT, Chang NT, Leonard WJ. Identification, cloning, and characterization of an immune activation gene. Proc Natl Acad Sci U S A. 1988; 85(24):9704-9708. (Biology). 查看参考
-
Napolitano M, Seamon KB, Leonard WJ. Identification of cell surface receptors for the Act-2 cytokine. J Exp Med. 1990; 172(1):285-289. (Biology). 查看参考
-
Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: Flow cytometry, IC/FCM Block). 查看参考
-
Raport CJ, Gosling J, Schweickart VL, Gray PW, Charo IF. Molecular cloning and functional characterization of a novel human CC chemokine receptor (CCR5) for RANTES, MIP-1beta, and MIP-1alpha. J Biol Chem. 1996; 271(29):17161-17166. (Biology). 查看参考
-
Samson M, Labbe O, Mollereau C, Vassart G, Parmentier M. Molecular cloning and functional expression of a new human CC-chemokine receptor gene. Biochemistry. 1996; 35(11):3362-3367. (Biology). 查看参考
-
Sherry B, Tekamp-Olson P, Gallegos C. one of those components, macrophage inflammatory protein 1 beta. J Exp Med. 1988; 168(6):2251-2259. (Biology). 查看参考
-
Ziegler SF, Tough TW, Franklin TL, Armitage RJ, Alderson MR. Induction of macrophage inflammatory protein-1 beta gene expression in human monocytes by lipopolysaccharide and IL-7. J Immunol. 1991; 147(7):2234-2239. (Biology). 查看参考
561120 Rev. 2
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.