Shade/CYP341a1/P450

Accession Number and Alternate nomenclature:  NP_996074;  Ecdysone 20-monooxygenase; CYP 341a1; CYPCCCXIVA1; CG13478.

Transposable elements are found in the genomes of nearly all eukaryotes. The recent completion of the Release 3 euchromatic genomic sequence of Drosophila melanogaster by the Berkeley Drosophila Genome Project has provided precise sequence for the repetitive elements in the Drosophila euchromatin. The genomic sequence to describe the euchromatic transposable elements in the sequenced strain of this species identified several new genes.  Out of 85 unknown and  8 novel family of transposable elements varying in copy number form 1-146 have been identified.  A total of 1500 full and partial transposable elements were identified that make up about 4% of the fruit fly genome. Most of these transposable elements reside outside genes (1). 

Spook or cytochrome P450a1 enzymes are involved in the detoxification of alkaloids and in xenobiotics resistance in insects.  Cytochrome P450s form a large and diverse family of heme-containing proteins capable of carrying out many different enzymatic reactions.  Some P450 proteins carry out reaction that are essential for process such as hormone synthesis, while others are P450 enzymes are involved in detoxification of environmental compounds.  Shade expression patterns in embryos and s stage of third instar larvae indicate the protein is in abundance in fat bodies, malpighian tubules and in distinct regions of the midgut, consistent with its role in detoxification (2). The biological functions also involve embryonic development via syncytical blastderm, ecdysone biosynthesis process, heat involution, cuticle development.   Shade gene sequence location is at 3L:5583181..5585677 is a 540 aa (65kDa) protein. There are at least 14 alleles are reported for this gene.      

The D. melanogaster Shade-selective antibodies were generated against unique antigenic sequence taken form the N-terminal region with in the amino acids 1-60 and from 175-225.  The sequence was conserved in several species of fruit flies, but was unique to shade/CYP341a1 protein and was not present in other proteins.  The Shade antibodies were affinity purified over immobilized antigen based chromatography, and the affinity purified immunoglobulins are stabilized in antibody stabilization buffer.  FabGennix Int. Inc., will provide limited quantities of antigenic blocking peptide for Shade antibodies.  FabGennix has a vast collection of antibodies to fruit fly and other insect proteins.  For a complete list of antibodies please visit www.FabGennix.com.  FabGennix Inc. will conjugate this and all its catalog antibodies with secondary enzymes (alk-Pase or HRP) or fluorescent probes upon request at a nominal cost.  FabGennix Int. Inc., will also provide and antigenic blocking peptide and western blot positive controls for its antibodies in ready-to-use buffer.

Immunogen:            Synthetic peptide taken form the Spook protein form within the N-terminal region (amino acid 51-60) and mid-region peptide (from with in aa 175-225), the peptides were post-synthetically modified and conjugated to carrier protein to achieve desired antigenicity.  The Shade peptides sequences are conserved various other specieis of fruit flys.

Concentration:         Shd-101AP and Shd-112AP:  IgG concentration 0.55-0.68mg/ml in antibody stabilization buffer.

Applications:           Antibody Shd-101AP and Shd-112AP are ideal for WB and IHC applications, other applications have not been tested.  This antibody does not cross react to other fly proteins. The species cross reactivity for this antibody is not examined fully.  The dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions for specific assay.  WB: > 1:500; IMM & i.p pull-down assays: n.d; IHC n.d.  ELISA <1:10,000; n.d does not necessary means antibody will not work in that applications.  Investigators using this antibody in applications not listed here can request for a complimentary sample. 

Reactivity:               This antibody detects a single band of approximately 65kDa in PC-Spk samples.  n.d does not necessarily means antibody is not suitable for that application, it simply indicate it has not been tried for that application. Investigators using this antibody in applications not listed here can ask for a free complimentary sample of this antibody.

Protocols:                Standard protocol for various applications (WB; IMM and IHC) of this antibody can be requested by calling Technical support line, basic information on this antibody is provided with the product specification sheet, however, FabGennix Inc.  strongly recommends investigators to optimize conditions for use of this antibody in their laboratories.  

Form/Storage:         The affinity-purified antibodies are supplied as stabilized product.  Store at -20^oC for long-term storage.  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 0.45u filter after every use for long-term storage.

For research use only, not for diagnostic or therapeutic use.