Merck

A1559

Sigma-Aldrich

Monoclonal Anti-Biotin–Agarose antibody produced in mouse

clone BN-34, purified immunoglobulin, PBS suspension

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Sinónimos:
Monoclonal Anti-Biotin
NACRES:
NA.46

origen biológico

mouse

Nivel de calidad

recombinante

expressed in mouse cell line

conjugado

agarose conjugate

forma del anticuerpo

purified immunoglobulin

antibody product type

primary antibodies

clon

BN-34, monoclonal

formulario

PBS suspension

extensión del etiquetado

2 mg antibody per mL resin

technique(s)

ELISA: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotipo

IgG1

capacidad

0.15-0.3 μmol/mL, agarose binding capacity (biotin-ligand conjugate)

application(s)

research pathology

Condiciones de envío

wet ice

temp. de almacenamiento

2-8°C

target post-translational modification

unmodified

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Este artículo
C5585B7653SAB4200680
recombinant

expressed in mouse cell line

recombinant

-

recombinant

-

recombinant

-

conjugate

agarose conjugate

conjugate

CY3 conjugate

conjugate

unconjugated

conjugate

-

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

antibody form

ascites fluid

antibody form

purified immunoglobulin

clone

BN-34, monoclonal

clone

BN-34, monoclonal

clone

BN-34, monoclonal

clone

BN-34, monoclonal

form

PBS suspension

form

buffered aqueous solution

form

-

form

buffered aqueous solution

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Descripción general

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.

Inmunógeno

Biotin-KLH conjugate

Aplicación

Applications in which this antibody has been used successfully, and the associated peer-reviewed papers, are given below.
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.
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.

Forma física

Suspension in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide

Nota de preparación

Prepared using purified antibody coupled to cyanogen bromide activated agarose

Otras notas

Polyclonal anti-biotin antibodies were shown to react with in vivo biotinylated proteins but not monoclonal anti-biotin antibodies.

Cláusula de descargo de responsabilidad

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.

Código de clase de almacenamiento

10 - Combustible liquids

Clase de riesgo para el agua (WGK)

WGK 3

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable


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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
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)
Protein phosphatase 1c associated with the cardiac sodium calcium exchanger 1 regulates its activity by dephosphorylating serine 68-phosphorylated phospholemman
Hafver T, et al.
The Journal of biological chemistry, 291(9), 4561-4579 (2016)
Alexander Brandis et al.
Journal of separation science, 40(16), 3358-3367 (2017-06-13)
A practical and relatively simple method to identify molecularly imprinted polymers capable of binding proteins via the molecular tagging (epitope-like) approach has been developed. In our two-step method, we first challenge a previously obtained anti-tag molecularly imprinted polymer with a

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