Fe superoxide dismutase

Accession # AAQ14560;  Alternate nomenclature: FeSOD

Kinetoplastid glycosomes contain a variety of metabolic activities, such as glycolysis, beta-oxidation of fatty acids, lipid biosynthesis, and purine salvage. Confinement and sequestration of all metabolic activities tend to protect other cellular components form oxidative damages caused by oxygen radicals produced during metabolism. In Leishmaniasis, an iron superoxide dismutase (FeSODB) is expressed in low levels in the early logarithmic promastigote stage and increases toward the stationary promastigote and amastigote stages. A second FeSODB2 enzymes was cloned from the same region but its expression was high in the early logarithmic promastigote stage and decreases toward the stationary promastigote and amastigote stages. Mutations in FeSODB1 results in significant reduction in parasite in human macrophages, suggesting that FeSODB1 plays an important role in parasite growth and survival by protecting glycosomes form superoxide toxicity (1). Other parasites like trypnasoma, overexpressing mitochondrial FeSOD (iron superoxide dismutase) were more resistant to the PCD stimulus, unambiguously indicating the participation of mitochondrial O2•− in the signaling process (2).      

The Leishmania FeSODB1 and FeSODB2 enzymes are cloned form L chagasi promastigote cDNA library using degenerate primers derived from conserved amino acids form the MnSOD and FeSOD, the two enzymes having high homology to FeSODB1 and FeSODB2 (3). Overexpression of these FeSODs in L. chagasi parasites also showed enhanced protection against the free radical generating agents, paraquat and nitroprusside.. FeSODB1 and B2 are highly homologous proteins with a MW of 25kDa (208aa). 

The FeSODB1/2-selective antibodies were generated against FeSOD protein.  The FeSOD1/2-selective antibodies are affinity purified against immobilized antigen based affinity chromatography which yielded high affinity epitope-specific antibodies.  The FeSOD1 antibodies label a protein of approximately 26kDa in Western blot using Western blot positive control for FeSOD1 (PC-FeSODB1).  FeSODB1/2-selective antibodies are also available in FITC conjugated preparations for confocal and immunocytochemical analyses.  FabGennix Inc. will also conjugate antibodies with many fluorescent probes upon request at a nominal charge.  Limited quantities of antigenic blocking peptide and western blot positive controls are also available (inquire before ordering).   FabGennix Inc. also provides antibodies to other targets of insect and parasite proteins, for a complete listing please visit www.FabGennix.com. 

Immunogen:            Purified peptide from FeSODB1 protein was covalently modified to carrier protein and emulsified in adjuvant before injected in to rabbits to make anti-FeSODB1 antibodies. There is a significant homology in the amino acid sequence of FeSODB enzymes that this antibody will label FeSODB1 in various species.                

Concentration:  Affinity purified FeSODB1-101AP antibody (0.5-0.6mg/ml) was stabilized in antibody stabilization buffer for long-term storage.

Applications:  FeSODB1-101AP antibody was tested in ELISA and western blotting applications at 1:500 dilution using PC-FeSODB1 samples.  Antibody dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions.  Application of this antibody in other protocols has not yet standardized. N.d  means applications for this antibody has not been tested but not necessarily means it cannot be used for that application.  WB: > 1:500; IMM & i.p pull-down assays: n.d; IHC n.d ().                            

Reactivity: The FeSODB1-101AP labels a 25kDa FeSODB1 protein in PC-FeSODB1 samples.

Protocols:  Standard protocol for various applications (WB, IMM, IHC) of this antibody can be obtained upon request. The specification sheet for FeSODB1 antibody will be supplied with each product.  FabGennix Inc., strongly recommends investigators to optimize conditions for use of this antibody in their laboratories.  

Storage:   The antiserum is supplied in antibody stabilization buffer with 0.02% azide.  The purified antibodies are isolated from immobilized antigen affinity column.  The affinity pure antibodies are supplied as 0.5-1 mg/ml IgG in antibody stabilization buffer.   For long-term storage of keep at -20^oC.  FabGennix Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi use antibody dilution buffer (Cat # DiluOBuffer).  Working solutions of antibodies in DiluOBuffer should be filtered through 0.45¿Ý filter after every use for long-term storage.

Notes:              Briefly centrifuge to collect liquid, heat or boil PC-FeSODB1tube for 1-2 minutes to dissolve any precipitate before use.  This product is “ready-to-use” for electrophoresis. After thawing store at room temperature, Repeated freezing and thawing may result in appearance of higher MW immunoreactive bands.   

New Reagents:  Now you can recycle your western blots (nitrocellulose, supported membranes and PVFD membranes) by using our StripObuffer (Cat FGI-1989).  Each stripping is guaranteed to give better signal (up to 8 stripping).  No strong pungent smell of reducing agents or heating required.  Block in 5X diluObuffer and you are ready for blotting with a new antibody