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Anti-Human B-Cell Activating Factor (BAFF)
Purified No Carrier Protein
(TNFSF13B, TALL-1, BLyS, THANK, zTNF4, BAFF, CD257, TNFSF20, Delta BAFF)
Monoclonal Antibody
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Prod. No.:B570
Clone:137314
Isotype:Mouse IgG1
Pkg. Size:500 µg, 1 mg
Storage:-20°C to -70°C Detailed storage instructions below.
Description
Specificity:
Mouse Anti-Human B-Cell Activating Factor (BAFF) (Clone 137314) recognizes an epitope on Human BAFF. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Background:
B-cell activating factor (BAFF) is the tumor necrosis factor ligand superfamily member 13B (TNFLSF13B), also known as B Lymphocyte Stimulator (BLyS) and TNF- and APOL-related leukocyte expressed ligand (TALL-1) and the Dendritic cell-derived TNF-like molecule (CD257 antigen; cluster of differentiation 257).

BAFF is a glycoprotein which undergoes glycosylation at residue 124. It is expressed as transmembrane protein on various cell types including monocytes, dendritic cells and bone marrow stromal cells. The transmembrane form can be cleaved from the membrane, generating a soluble protein fragment. BAFF is the natural ligand of three unusual tumor necrosis factor receptors named BAFF-R, TACI, and BCMA, all of which have differing binding affinities for it. These receptors are expressed mainly on mature B lymphocytes (TACI is also found on a subset of T-cells and BCMA on plasma cells). TACI binds worst since its affinity is higher for a protein similar to BAFF, called A Proliferation Inducing Ligand (APRIL). BCMA displays an intermediate binding phenotype and will work with either BAFF or APRIL to varying degrees. Signaling trough BAFF-R and BCMA stimulates B lymphocytes to undergo proliferation and to counter apoptosis. All these ligands act as heterotrimers (i.e. three of the same molecule) interacting with heterotrimeric receptors.

BAFF provides critical survival signals to a subset of B cells with intermediate maturation status (T2 B cells) during the immune response. BAFF also plays an important role in the development of lymphoid tissue and enhances the survival of activated memory B cells. Human and mouse BAFF share 86% aa sequence identity.

Known Species Cross Reactivity: Hu+
Host Species
Mouse
Immunogen
Purified Recombinant Human BAFF (>98%)
Formulation
This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.
Reconstitution
This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1 ml. This will result in a final antibody concentration of 500 µg/ml.
Endotoxin
<0.1 EU/µg as determined by the LAL method
Storage and Stability
The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.
Reported Applications
Indirect ELISA, FC, WB
Indirect ELISA: This antibody can be used in an indirect detection ELISA at 0.5-1.0 µg/ml with a suitable second step reagent such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114). The detection sensitivity of this indirect ELISA for Human BAFF is approximately 1 ng/well.
Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing BAFF. A suggested method would be to stain cells expressing BAFF with 0.25 µg per 1-5 x 105 cells in a total staining volume of ≤200 µl followed by Goat Anti-Mouse IgG (H&L)-R-phycoerythrin (Leinco Prod. No.: M1189).
Western Blotting: To detect Human BAFF this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for rhBAFF is approximately 25 ng/lane and 1 ng/lane under non-reducing and reducing conditions, respectively. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used.
Each investigator should determine their own optimal working dilution for specific applications.


Country of Origin
USA
Related Products
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Goat Anti-Mouse IgG (H&L)-R-phycoerythrin (Leinco Prod. No.: M1189)
TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343)
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