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Purified Mouse Anti-Human Thioredoxin
Purified Mouse Anti-Human Thioredoxin

Detection of Trx (1-104) by Western Blotting: Recombinant huTrx (100, 50 25 or 12.5 ng-lanes 1 -4, respectively) or dilutions of U937 cell lysate (5x10e4 or 2.5x10e4 cell equivalent-- lanes 5 and 6 respectively) were separated on 14% SDS- PAGE under reducing conditions. The separated proteins were transferred to PVDF membrane, and probed with Purified Mouse Anti-Human Thioredoxin (1 µg/ml, Cat. No. 559969). BD Pharmingen™ AKP Rat Anti-Mouse IgG1 (Cat. No. 557272) was used as a secondary antibody at 1:1000 dilution, and the blot was developed with BCIP/NBT substrate (Sigma). The 2G11 antibody detects a single, approx. 12 kDa band protein in lanes 1 - 4 (rhuTrx) as well as well as in the cellular lysate, that corresponds to the molecular weight of Trx (left panel).

Detection of Trx (1-104) by Western Blotting: Recombinant huTrx (100, 50 25 or 12.5 ng-lanes 1 -4, respectively) or dilutions of U937 cell lysate (5x10e4 or 2.5x10e4 cell equivalent-- lanes 5 and 6 respectively) were separated on 14% SDS- PAGE under reducing conditions. The separated proteins were transferred to PVDF membrane, and probed with Purified Mouse Anti-Human Thioredoxin (1 µg/ml, Cat. No. 559969). BD Pharmingen™ AKP Rat Anti-Mouse IgG1 (Cat. No. 557272) was used as a secondary antibody at 1:1000 dilution, and the blot was developed with BCIP/NBT substrate (Sigma). The 2G11 antibody detects a single, approx. 12 kDa band protein in lanes 1 - 4 (rhuTrx) as well as well as in the cellular lysate, that corresponds to the molecular weight of Trx (left panel).

Product Details
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BD Pharmingen™
TRX (1-104)
Human (QC Testing)
Mouse IgG1
Recombinant human Trx
Western blot (Routinely Tested), Intracellular staining (flow cytometry) (Tested During Development)
0.5 mg/ml
AB_398684
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

The 2G11 antibody is suitable to detect Trx (1-104) in western blots. 2G11 is useful for Intracellular Staining and Flow cytometry with an appropriate conjugated secondary antibody. A useful control for demonstrating specificity of staining with conjugated 2G11 is to pre-block the fixed/permeabilized cells with unlabeled 2G11 antibody (Cat. No. 559969) prior to staining.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. 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.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
559969 Rev. 2
Antibody Details
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2G11/TRX

The immunogen used to raise the 2G11 hybridoma was recombinant full length human thioredoxin (Trx) is an intracellular, multifunctional 12 kD (104 a.a.) protein with reduction/oxidation (redox) activity expressed by most cells of the human body. Trx is also secreted via the leaderless pathway (i.e. like IL-1β) by cells such as monocytes and lymphocytes upon activation or in response to oxidative stress. A truncated, 10 kD form of Trx (Trx1-84) has also been reported. Secreted Trx (also known as adult T-cell leukemia derived factor (ADF)) exhibits several cytokine- and chemokine-like functions such as activating eosinophils and granulocytes and chemotaxis for neutrophils, monocytes and T cells. 2G11 reacts exclusively with Trx(1-104), but not with its truncated form.

559969 Rev. 2
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
559969 Rev.2
Citations & References
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Development References (4)

  1. Bertini R, Howard OM, Dong HF, et al. Thioredoxin, a redox enzyme released in infection and inflammation, is a unique chemoattractant for neutrophils, monocytes, and T cells. J Exp Med. 1999; 189(11):1783-1789. (Biology). View Reference
  2. 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: IC/FCM Block). View Reference
  3. Sahaf B, Söderberg A, Spyrou G, et al. Thioredoxin expression and localization in human cell lines: detection of full-length and truncated species. Exp Cell Res. 1997; 236(1):181-192. (Biology). View Reference
  4. Yoshida S, Katoh T, Tetsuka T, Uno K, Matsui N, Okamoto T. Involvement of thioredoxin in rheumatoid arthritis: its costimulatory roles in the TNF-alpha-induced production of IL-6 and IL-8 from cultured synovial fibroblasts. J Immunol. 1999; 163(1):351-358. (Biology). View Reference
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559969 Rev. 2

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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.