antibody

Phosphatase, Alkaline, Escherichia coli (E. coli Alkaline Phosphatase, bacterial only)

P4071-01A

100 ul

monoclonal

mouse

nonconjugated

2Q2148

Phosphatase, Alkaline, Escherichia coli (E. coli Alkaline Phosphatase, bacterial only)

mouse

Mab

Antibodies

100 ul

2Q2148

IgG2a

Escherichia coli (E. coli) is a Gram negative bacterium that is commonly found in the lower intestine of warm-blooded organisms (endotherms). Most E. coli strains are harmless, but some, such as serotype O157:H7, can cause serious food poisoning in humans, and are occasionally responsible for costly product recalls.[1][2] The harmless strains are part of the normal flora of the gut, and can benefit their hosts by producing vitamin K2,[3] or by preventing the establishment of pathogenic bacteria within the intestine.[4][5] E. coli are not always confined to the intestine, and their ability to survive for brief periods outside the body makes them an ideal indicator organism to test environmental samples for fecal contamination.[6][7] The bacteria can also be grown easily and its genetics are comparatively simple and easily-manipulated or duplicated through a process of metagenics, making it one of the best-studied prokaryotic model organisms, and an important species in biotechnology and microbiology. E. coli was discovered by German pediatrician and bacteriologist Theodor Escherich in 1885,[6] and is now classified as part of the Enterobacteriaceae family of gamma-proteobacteria.[8] A strain of E. coli is a sub-group within the species that has unique characteristics that distinguish it from other E. coli strains. These differences are often detectable only on the molecular level; however, they may result in changes to the physiology or lifecycle of the bacterium. For example, a strain may gain pathogenic capacity, the ability to use a unique carbon source, the ability to inhabit a particular ecological niche or the ability to resist antimicrobial agents. Different strains of E. coli are often host-specific, making it possible to determine the source of fecal contamination in environmental samples.[6][7] For example, knowing which E. coli strains are present in a water sample allows to make assumptions about whether the contamination originated from a human, another mammal or a bird. Escherichia coli cells propel themselves with flagella (long, thin structures) arranged as bundles that rotate counter-clockwise, generating torque to rotate the bacterium clockwise. E. coli is Gram-negative, facultative anaerobic and non-sporulating. Cells are typically rod-shaped and are about 2 micrometres (um) long and 0.5 um in diameter, with a cell volume of 0.6-0.7cm3.[9] It can live on a wide variety of substrates. E. coli uses mixed-acid fermentation in anaerobic conditions, producing lactate, succinate, ethanol, acetate and carbon dioxide. Since many pathways in mixed-acid fermentation produce hydrogen gas, these pathways require the levels of hydrogen to be low, as is the case when E. coli lives together with hydrogen-consuming organisms such as methanogens or sulfate-reducing bacteria.[10] Optimal growth of E. coli occurs at 37°C but some laboratory strains can multiply at temperatures of up to 49°C.[11] Growth can be driven by aerobic or anaerobic respiration, using a large variety of redox pairs, including the oxidation of pyruvic acid, formic acid, hydrogen and amino acids, and the reduction of substrates such as oxygen, nitrate, dimethyl sulfoxide and trimethylamine N-oxide.[12] Strains that possess flagella can swim and are motile. The flagella have a peritrichous arrangement.[13] E. coli and related bacteria possess the ability to transfer DNA via bacterial conjugation, transduction or transformation, which allows genetic material to spread horizontally through an existing population. This process led to the spread of the gene encoding shiga toxin from Shigella to E. coli O157:H7, carried by a bacteriophage.[14] Applications: Suitable for use in Western Blot, Immunoprecipitation, Immunocytochemistry. Other applications not tested. Recommended Dilution: Western Blot: 1:5000. Optimal dilutions to be determined by the researcher. Assay: Preparation of E. coli TnphoA transformants: E. coli strain CC118 was transformed with plasmid pGEM-3Z containing TnphoA insertional mutations in the p101 gene of Mycoplasma hyorhinis, which encodes a protein with a typical N-terminal prokaryotic single peptide (5). Identification of fusion protein: Transformants are grown in 2XYT medium to OD600=0.6. Cells were centrifuged 3 minutes at 10,000 x g, suspended in SDS-PAGE sample buffer, heated at 100°C for 5 minutes, frozen and thawed and centrifuged as above at RT to remove insoluble material. The sample is applied at 9% to a SDS-PAGE gel, and Western immunoblot is performed as described (5). Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Ec

E. coli Alkaline Phosphatase

Recognizes E. coli Alkaline Phosphatase. This antibody has a high affinity and recognizes an AP determinant resistant to denaturation by SDS-PAGE. It is therefore ideally suited for sensitive and specific detection of AP fusion proteins by Western blot analysis of E. coli transformants expressing fusion products.

Ascites

Supplied as a liquid. No preservative added.

Not determined

Product Type: Mab Isotype: IgG2a Clone No: 2Q2148 Host: mouse Source: E. coli Concentration: Not determined Form: Supplied as a liquid. No preservative added. Purity: Ascites Immunogen: E. coli Alkaline Phosphatase Specificity: Recognizes E. coli Alkaline Phosphatase. This antibody has a high affinity and recognizes an AP determinant resistant to denaturation by SDS-PAGE. It is therefore ideally suited for sensitive and specific detection of AP fusion proteins by Western blot analysis of E. coli transformants expressing fusion products. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

IC IP WB

-20°C

1. PNAS.USA (1985) 82:5107–5111. 2. PNAS.USA (1985) 82:8129–8133. 3. Science (1986) 233:1403–1408. 4. J. Bacteriol. (1990) 172:515–518. 5. J. Bacteriol. (1991) 173:2035–2044. General References: 1. "Escherichia coli O157:H7". CDC Division of Bacterial and Mycotic Diseases. Retrieved on 2007-01-25. 2. Vogt RL, Dippold L (2005). "Escherichia coli O157:H7 outbreak associated with consumption of ground beef, June-July 2002". Public Health Rep 120 (2): 174–8. PMID 15842119. 3. Bentley R, Meganathan R (01 September 1982). "Biosynthesis of vitamin K (menaquinone) in bacteria". Microbiol. Rev. 46 (3): 241–80. PMID 6127606. PMC: 281544. 4. Hudault S, Guignot J, Servin AL (July 2001). "Escherichia coli strains colonising the gastrointestinal tract protect germfree mice against Salmonella typhimurium infection". Gut 49 (1): 47–55. doi:10.1136/gut.49.1.47. PMID 11413110. 5. Reid G, Howard J, Gan BS (September 2001). "Can bacterial interference prevent infection?". Trends Microbiol. 9 (9): 424–8. doi:10.1016/S0966-842X(01)02132-1. PMID 11553454. 6. Feng P, Weagant S, Grant, M (2002-09-01). "Enumeration of Escherichia coli and the Coliform Bacteria". Bacteriological Analytical Manual (8th ed.). FDA/Center for Food Safety & Applied Nutrition. Retrieved on 2007-01-25. 7. Thompson, Andrea (2007-06-04). "E. coli Thrives in Beach Sands". Live Science. Retrieved on 2007-12-03. 8. "Escherichia". Taxonomy Browser. NCBI. Retrieved on 2007-11-30. 9. Kubitschek HE (01 January 1990). "Cell volume increase in Escherichia coli after shifts to richer media". J. Bacteriol. 172 (1): 94–101. PMID 2403552. PMC: 208405. 10. Madigan MT, Martinko JM (2006). Brock Biology of microorganisms (11th ed.). Pearson. ISBN 0-13-196893-9. 11. Fotadar U, Zaveloff P, Terracio L (2005). "Growth of Escherichia coli at elevated temperatures". J. Basic Microbiol. 45 (5): 403–4. doi:10.1002/jobm.200410542. PMID 16187264. 12. Ingledew WJ, Poole RK (1984). "The respiratory chains of Escherichia coli". Microbiol. Rev. 48 (3): 222–71. PMID 6387427. 13. Darnton NC, Turner L, Rojevsky S, Berg HC, On torque and tumbling in swimming Escherichia coli. J Bacteriol. 2007 Mar;18 Brüssow H, Canchaya C, Hardt WD (September 2004). "Phages and the evolution of bacterial pathogens: from genomic rearrangements to lysogenic conversion". Microbiol. Mol. Biol. Rev. 68 (3): 560–602. doi:10.1128/MMBR.68.3.560-602.2004. PMID 15353570. PMC: 515249.