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Purified Mouse Anti-FKBP-12
Purified Mouse Anti-FKBP-12

Western blot analysis of FKBP-12 expression on Jurkat cell lysate. Jurkat cell lysate (ATCC TIB-152) was stained with Purified Mouse Anti-FKBP-12 (Cat. No. 554091) at 2, 1, 0.5, and 0.25 µg/ml (lane 1, 2, 3, 4, respectively), then visualized with HRP Goat Anti-Mouse Ig (Cat. No. 554002).  FKBP-12 is identified by 12 kDa band.

Western blot analysis of FKBP-12 expression on Jurkat cell lysate. Jurkat cell lysate (ATCC TIB-152) was stained with Purified Mouse Anti-FKBP-12 (Cat. No. 554091) at 2, 1, 0.5, and 0.25 µg/ml (lane 1, 2, 3, 4, respectively), then visualized with HRP Goat Anti-Mouse Ig (Cat. No. 554002).  FKBP-12 is identified by 12 kDa band.

Product Details
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BD Pharmingen™
Human (QC Testing), Mouse, Rat (Reported)
Mouse IgG1
Human Recombinant FKBP-12
Western blot (Routinely Tested), Immunohistochemistry-paraffin (Reported)
12 kDa
0.5 mg/ml
AB_2102852
Aqueous buffered solution containing ≤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.

Recommended Assay Procedures

Applications include western blot analysis, immunohistochemistry of cytospins (reported) and of paraffin-embedded tissue sections. Jurkat T cells (ATCC TIB-152) are suggested as a positive control.

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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
554091 Rev. 13
Antibody Details
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2C1-87

FKBP-12 (for FK506 binding protein-12 kDa) is a ubiquitously expressed protein involved in T-cell signaling that binds to FK506. FK506 belongs to a class of proteins that have peptidyl-prolyl cis-trans isomerase (PPIase) activity. This class of proteins is thought to be involved in folding proteins and peptides into their native conformations. It is thought that peptidyl-prolyl isomerization may play a role in regulating components of the T-cell signal transduction pathway such as transcription factors, protein kinase activation, and ion channel function. FK506, like cyclosporin, is an agent that inhibits T- and B-cell early activation processes that are key to an effective immune response. In T cells these agents disrupt a step in the signal transduction pathway from the T-cell receptor to cytokine genes involved in coordinating the immune response. In particular, they seem to disrupt pathways that induce a measurable rise in Ca2+. NFAT, a  transcription factor essential for early T-cell activation, appears to be a specific target of FK506 and cyclosporin because transcription directed by NF-AT is specifically blocked in T cells treated with these agents. NFAT is formed when a signal from the T-cell antigen receptor induces a pre-existing cytoplasmic subunit to translocate to the nucleus and combine with a newly synthesized nuclear subunit of NFAT. When FK506 binds to FKBP-12, the complex blocks translocation of the cytoplasmic subunit of NF-AT into the nucleus by inhibiting a Ca2+-dependent phosphatase. Since the cytoplasmic subunit is not translocated into the nuclear, the formation of NFAT and its subsequent transcriptional  activity is blocked. The reduced molecular weight of FKBP-12 is 12 kDa.

The 2C1-87 antibody recognizes FKBP-12. Specifically it has been shown to recognize human, mouse, and rat FKBP-12. Recombinant FKBP-12 was used as immunogen. The antibody was originally characterized by biochemical, immunohistochemical, and functional assays.

554091 Rev. 13
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
554091 Rev.13
Citations & References
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Development References (3)

  1. Flanagan WM, Corthésy B, Bram RJ, Crabtree GR. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A. Nature. 1991; 352(6338):803-807. (Biology). View Reference
  2. Kobayashi M, Ohtsuka K, Tamura K, et al. Production of monoclonal antibody against recombinant human FKBP-12 and subcellular localization of FKBP-12 in human mononuclear and polymorphonuclear cells. Transplant Proc. 1993; 125(1):655-657. (Immunogen: Immunohistochemistry, Western blot). View Reference
  3. Siekierka JJ, Wiederrecht G, Greulich H, et al. The cytosolic-binding protein for the immunosuppressant FK-506 is both a ubiquitous and highly conserved peptidyl-prolyl cis-trans isomerase. J Biol Chem. 1990; 265(34):21011-21015. (Clone-specific: Functional assay, Immunohistochemistry). View Reference
554091 Rev. 13

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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.