MAB1012
Anti-Alkaline Phosphatase Antibody, E. coli, bacterial only
ascites fluid, Chemicon®
Synonym(s):
AP, Alk. Phos.
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
mouse
Quality Level
antibody form
ascites fluid
antibody product type
primary antibodies
clone
monoclonal
species reactivity
E. coli
manufacturer/tradename
Chemicon®
technique(s)
immunocytochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG2a
UniProt accession no.
shipped in
dry ice
target post-translational modification
unmodified
Gene Information
Escherichia coli ... PhoA(945041)
Specificity
Alkaline phosphatase (AP). MAB1012 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.
Immunogen
Epitope: E. coli, bacterial only
F201C1B
Application
Anti-Alkaline Phosphatase Antibody, E. coli, bacterial only detects level of Alkaline Phosphatase & has been published & validated for use in IP, WB & IC.
Western blot at 1:5,000. 42-45kDa on SDS-PAGE,reducing gels for natural E. coli alkaline phosphatase monomer. Alkaline phosphatase fusion proteins will vary depending upon the target fusion protein.
Immunocytochemistry: reacts with E.coli. AP fusion protein targets in acetone fixed cell preparations. 1:4000, other fixatives or conditions untested.
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 (Yogev et al. 1991).
Identification of fusion protein with MAB1012 transformants: 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 room temperature to remove insoluble material. The sample is applied at 9% to a SDS-PAGE gel, and Western immunoblot is performed as described (Yogev et al. 1991).
Immunoprecipitation: 5μL of antibody per 500μL of lysate in RIPA or 0.5% triton X-100 solutions.
Optimal working dilutions must be determined by end user.
Immunocytochemistry: reacts with E.coli. AP fusion protein targets in acetone fixed cell preparations. 1:4000, other fixatives or conditions untested.
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 (Yogev et al. 1991).
Identification of fusion protein with MAB1012 transformants: 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 room temperature to remove insoluble material. The sample is applied at 9% to a SDS-PAGE gel, and Western immunoblot is performed as described (Yogev et al. 1991).
Immunoprecipitation: 5μL of antibody per 500μL of lysate in RIPA or 0.5% triton X-100 solutions.
Optimal working dilutions must be determined by end user.
Physical form
Ascites. Contains no preservative.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
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Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Find documentation for the products that you have recently purchased in the Document Library.
Sequence and TnphoA analysis of a Mycoplasma hyorhinis protein with membrane export function.
Yogev, D, et al.
Journal of Bacteriology, 173, 2035-2044 (1991)
A genetic approach to analyzing membrane protein topology.
Manoil, C and Beckwith, J
Science (New York, N.Y.), 233, 1403-1408 (1986)
Both the stroma and thylakoid lumen of tobacco chloroplasts are competent for the formation of disulphide bonds in recombinant proteins.
Julia Bally,Eric Paget,Michel Droux,Claudette Job,Dominique Job,Manuel Dubald
Plant Biotechnology Journal null
Fusions of secreted proteins to alkaline phosphatase: an approach for studying protein secretion.
Hoffman, C S and Wright, A
Proceedings of the National Academy of Sciences of the USA, 82, 5107-5111 (1985)
TnphoA: a transposon probe for protein export signals.
Manoil, C and Beckwith, J
Proceedings of the National Academy of Sciences of the USA, 82, 8129-8133 (1985)
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