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Biotin Mouse anti-Human Blys (BAFF)
Biotin Mouse anti-Human Blys (BAFF)

Human Blys sandwich ELISA curve. The curve was generated by a sandwich ELISA using Purifed Mouse anti-Human Blys (BAFF, H32-406, Cat. No. 558289) as capture antibody, doubling dilutions of recombinant human Blys protein and Biotin Mouse anti-Human Blys (BAFF, Cat. No. 558290) as detection antibody. Streptavidin-HRP (Cat. No. 554066) plus TMB substrate (Cat. No. 555214) were used to develop the ELISA.

Human Blys sandwich ELISA curve. The curve was generated by a sandwich ELISA using Purifed Mouse anti-Human Blys (BAFF, H32-406, Cat. No. 558289) as capture antibody, doubling dilutions of recombinant human Blys protein and Biotin Mouse anti-Human Blys (BAFF, Cat. No. 558290) as detection antibody. Streptavidin-HRP (Cat. No. 554066) plus TMB substrate (Cat. No. 555214) were used to develop the ELISA.

Product Details
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BD Pharmingen™
Mouse IgG1, κ
ELISA (Routinely Tested)
0.5 mg/ml
AB_647168
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.

Recommended Assay Procedures

The biotinylated H32-544 mAb is useful as a detection antibody for a sandwich ELISA that measures human Blys protein when paired with Purified Mouse anti-Human Blys (Cat. No. 558289) as capture antibody and recombinant human Blys protein as standard. The biotinylated H32-544 antibody should be titrated between 0.25 - 2 µg/ml to determine its optimal concentration. To obtain linear standard curves, doubling dilutions of recombinant human Blys protein ranging from 1.56 - 100 ng/ml are recommended. For specific methodology measuring the levels of cytokines, chemokines, and inflammatory mediators and their receptors, see protocol "Cytokine ELISA" under "ELISA and ELISPOT" at http://www.bdbiosciences.com/us/s/resources. For this ELISA antibody pair, no cross-reactivity has been observed with the following: human TNF, TNFRI and TNFRII. This ELISA antibody pair shows a limited cross-reactivity with mouse Blys.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558290 Rev. 2
Antibody Details
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H32-544

Antibody H32-544 reacts with B-lymphocyte stimulator protein (Blys), also known as BAFF (B cell activating factor), is a member of the TNF ligand superfamily. Members of the TNF receptor and ligand superfamily play a critical role in regulating immune responses. Blys is a type II membrane protein expressed on cells of myeloid lineage including monocytes, dendritic cells, and activated T cells. Blys exists in both a membrane-bound and soluble form. In its soluble form, Blys is a potent B cell  growth factor in costimulation assays, suggesting a role in monocyte-driven B cell activation. Blys has also been linked to B cell hyperplasia and autoimmunity.

558290 Rev. 2
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
558290 Rev.2
Citations & References
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Development References (4)

  1. Craxton A, Magaletti D, Ryan EJ, and Clark EA. Macrophage- and dendritic cell-dependent regulation of human B-cell proliferation requires the TNF family ligand BAFF. Blood. 2002; 101:4464-4471. (Biology).
  2. Mackay F, Sarosi I, Xia X-Z, et al. Severe B cell hyperplasia and autoimmune disease in TALL-1 transgenic mice. Proc Natl Acad Sci U S A. 2000; 97:3370-3375. (Biology).
  3. Mackay F, Shcneider P, Rennert P, and Browing J. BAFF and APRIL: a tutorial on B cell survival. Annu Rev Immunol. 2003; 21:231-264. (Biology: ELISA).
  4. Moore PA, Belvedere O, Orr A, et al. BlyS: member of the tumor necrosis factor family and B lymphocyte stimulator. Science. 1999; 285:260-263. (Biology).
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558290 Rev. 2

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