A1559
Monoclonal Anti-Biotin–Agarose antibody produced in mouse
clone BN-34, purified immunoglobulin, PBS suspension
Synonym(s):
Monoclonal Anti-Biotin
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About This Item
UNSPSC Code:
12352203
NACRES:
NA.46
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biological source
mouse
recombinant
expressed in mouse cell line
conjugate
agarose conjugate
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
BN-34, monoclonal
form
PBS suspension
extent of labeling
2 mg antibody per mL resin
technique(s)
ELISA: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG1
capacity
0.15-0.3 μmol/mL, agarose binding capacity (biotin-ligand conjugate)
application(s)
research pathology
shipped in
wet ice
storage temp.
2-8°C
target post-translational modification
unmodified
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General description
Biotin is an essential vitamin required for cells in living organisms and in cultures. The high binding affinity of biotin to egg white avidin or bacteria-derived streptavidin has been used to design immunoassays for detecting antigen-antibody interactions. The use of monoclonal anti-biotin IgG can enhance the sensitivity of such avidin-biotin immunoassays. The monoclonal antibody functions by selectively enlarging and bridging the avidin-biotin-enzyme complexes, thereby increasing the signals from substrate conversion. Furthermore, this antibody can be used in many other applications where biotin may be introduced as target label.
The antibody recognizes free biotin and biocytin using competitive ELISA. Using indirect ELISA or immunohistochemical staining, the antibody also recognizes biotin conjugated to various immunoglobulins. Specificity is evaluated using biotinylated goat antibodies to human and rabbit antigens coated on multiwell plates.
Immunogen
Biotin-KLH conjugate
Application
Applications in which this antibody has been used successfully, and the associated peer-reviewed papers, are given below.
Chromatin immunoprecipitation (1 paper)
Chromatin immunoprecipitation (1 paper)
Monoclonal Anti-Biotin-Agarose antibody produced in mouse has been used in:
- pulldown assay
- coimmunoprecipitation
- chromatin immunoprecipitation
- purification of biotinylated compounds and biotin-containing compounds
Mouse monoclonal anti-biotin antibody was used to identify by western blot S-nitrosylated proteins that had been labeled with a biotin moeity based on the biotin-switch method. No reactivity was visible on the blots when the biotin switch method was inhibited by treatment with 100 mM dithiothreitol showing that this antibody shows no unspecific cross reactivity. The mouse monoclonal anti-biotin antibody does cross react with Arabidopsis plant by western blot, no cross reactivity with unbiotinylated proteins.
In some applications, localization of biotinylated probes with avidin produces high background levels. Anti-biotin reagents may be substituted for avidin to decrease non-specific binding.
In some applications, localization of biotinylated probes with avidin produces high background levels. Anti-biotin reagents may be substituted for avidin to decrease non-specific binding.
The antibody may be used in amplification techniques, ELISA, immunoblotting, in situ nucleic acid hybridization, flow cytometry, image analysis or confocal microscopy.
In some applications, localization of biotinylated probes with avidin produces high background levels. Anti-biotin reagents may be substituted for avidin to decrease non-specific binding.
In some applications, localization of biotinylated probes with avidin produces high background levels. Anti-biotin reagents may be substituted for avidin to decrease non-specific binding.
Physical form
Suspension in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide
Preparation Note
Prepared using purified antibody coupled to cyanogen bromide activated agarose
Other Notes
Polyclonal anti-biotin antibodies were shown to react with in vivo biotinylated proteins but not monoclonal anti-biotin antibodies.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Storage Class Code
10 - Combustible liquids
WGK
WGK 3
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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Emmanouil Rampakakis et al.
Nucleic acids research, 37(17), 5714-5724 (2009-07-30)
Initiation of DNA replication involves the ordered assembly of the multi-protein pre-replicative complex (pre-RC) during G(1) phase. Previously, DNA topoisomerase II (topo II) was shown to associate with the DNA replication origin located in the lamin B2 gene locus in
A two-step strategy to visually identify molecularly imprinted polymers for tagged proteins
Brandis A, et al.
Journal of Separation Science, 40(16), 3358-3367 (2017)
F J Germino et al.
Proceedings of the National Academy of Sciences of the United States of America, 90(3), 933-937 (1993-02-01)
We describe an in vivo approach for the isolation of proteins interacting with a protein of interest. The protein of interest is "tagged" with a portion of the biotin carboxylase carrier protein (BCCP), encoded on a specially constructed plasmid, so
Transient dsDNA breaks during pre-replication complex assembly
Rampakakis E and Zannis-Hadjopoulos M
Nucleic Acids Research, 37(17), 5714-5724 (2009)
Namrata D Udeshi et al.
Nature methods, 14(12), 1167-1170 (2017-10-19)
Although purification of biotinylated molecules is highly efficient, identifying specific sites of biotinylation remains challenging. We show that anti-biotin antibodies enable unprecedented enrichment of biotinylated peptides from complex peptide mixtures. Live-cell proximity labeling using APEX peroxidase followed by anti-biotin enrichment
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Pull-down assays, reagents, and protocols for investigating in vitro protein-protein interactions using affinity or GST pull-down, tandem affinity purification (TAP), and co-immunoprecipitation methods.
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