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A8592

ANTI-FLAG® M2-Peroxidase (HRP) antibody, Mouse monoclonal

clone M2, purified immunoglobulin, buffered aqueous glycerol solution

ANTI-FLAG® M2-Peroxidase (HRP) antibody, Mouse monoclonal

Synonym(s):

Monoclonal ANTI-FLAG® M2, Anti-ddddk, Anti-dykddddk

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About This Item

NACRES:
NA.32
UNSPSC Code:
41106514
Conjugate:
peroxidase conjugate
Clone:
M2, monoclonal
Application:
ELISA (i)
Citations:
1904
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biological source

mouse

Quality Level

conjugate

peroxidase conjugate

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

M2, monoclonal

form

buffered aqueous glycerol solution

species reactivity

all

concentration

~1 mg/mL

technique(s)

indirect ELISA: 1:20,000

isotype

IgG1

immunogen sequence

DYKDDDDK

shipped in

wet ice

storage temp.

−20°C

Gene Information

human ... GPX1(2876)

General description

The Monoclonal ANTI-FLAG M2-Peroxidase is a mouse IgG antibody covalently conjugated to horseradish peroxidase ( HRP). The antibody binds to FLAG fusion proteins and recognizes the FLAG epitope at N-terminal, Met-N-terminal, C-terminal, and internal FLAG peptides.

Application

For simple, one-step detection by immunocytochemistry, ELISA, or Western blotting. Especially useful in detection of FLAG fusion proteins expressed in murine host, where secondary anti-mouse antibodies may cause cross-reactivity.
Suggested dilution for immunocytochemistry and western blotting 1:100 to 1:1000
Suggested dilution for ELISA 1:20,000

Learn more product details in our FLAG® application portal.

Physical form

Solution in phosphate buffered saline containing 50% glycerol plus preservative and stabilizer

Preparation Note

Dilute ANTI-FLAG M2-Peroxidase solution in Tris Buffered Saline (TBS): 0.05 M Tris, pH 7.4, with 0.15 M NaCl.

Legal Information

ANTI-FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany

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This Item
A9469F3040SAB4301135
species reactivity

all

species reactivity

all

species reactivity

all

species reactivity

all

clone

M2, monoclonal

clone

M2, monoclonal

clone

M1, monoclonal

clone

polyclonal

biological source

mouse

biological source

mouse

biological source

(Bioreactor)

biological source

rabbit

conjugate

peroxidase conjugate

conjugate

alkaline phosphatase conjugate

conjugate

unconjugated

conjugate

unconjugated

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

antibody form

affinity isolated antibody

technique(s)

indirect ELISA: 1:20,000

technique(s)

indirect ELISA: 1:20,000

technique(s)

western blot: 10 μg/mL

technique(s)

western blot: 1:1000-1:5000 (Cell Lysate)


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signalword

Danger

Hazard Classifications

Aquatic Acute 1 - Aquatic Chronic 1 - Eye Dam. 1 - Skin Corr. 1C - Skin Sens. 1

Storage Class

8A - Combustible corrosive hazardous materials

flash_point_f

Not applicable

flash_point_c

Not applicable



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Related Content


Linda Brunotte et al.
Journal of virology, 85(1), 324-333 (2010-10-29)
The 200-kDa L protein of arenaviruses plays a central role in viral genome replication and transcription. This study aimed at providing evidence for the domain structure of L protein by combining bioinformatics with a stepwise mutagenesis approach using the Lassa
Mickaël Lelek et al.
Nature communications, 6, 6483-6483 (2015-03-07)
The molecular mechanisms that allow HIV to integrate into particular sites of the host genome are poorly understood. Here we tested if the nuclear pore complex (NPC) facilitates the targeting of HIV integration by acting on chromatin topology. We show
Martin Roth et al.
Development (Cambridge, England), 137(9), 1553-1562 (2010-04-02)
FoxG1 is a conserved transcriptional repressor that plays a key role in the specification, proliferation and differentiation of the telencephalon, and is expressed from the earliest stages of telencephalic development through to the adult. How the interaction with co-factors might



Global Trade Item Number

SKUGTIN
A8592-5X1MG04061833396889
A8592-.2MG04061835511761
A8592-1MG04061833392409

Questions

1–7 of 7 Questions  
  1. I am currently using your product A8592 HRP in my analytical method with QuantaBlu fluorescence substrate complex. Is it possible to use TMB substrate instead of QuantaBlu fluorescence substrate with the A8592 HRP conjugate?

    1 answer
    1. A8592 is an HRP-conjugated antibody. The product information sheet can be downloaded here:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/111/306/a8592dat-mk.pdf

      Although we do not routinely test 3,3′,5,5′-tetramethylbenzidine (TMB) in-house, it is a standard chromogenic substrate for horseradish peroxidase, so it is expected to work with the A8592 conjugate.

      QuantaBlu from ThermoFisher is a fluorogenic substrate; TMB is colorimetric.
      You should expect lower sensitivity with TMB compared with QuantaBlu, as fluorescence detection generally offers higher sensitivity.
      The TMB-HRP reaction produces a soluble end-product that is blue in color and can be read spectrophotometrically at 370 nm or 650 nm.
      The reaction may be stopped with acid, resulting in a yellow solution that is read at 450 nm.

      Published protocol for use of A8592 together with TMB include:

      https://pmc.ncbi.nlm.nih.gov/articles/PMC3709174/pdf/cpb-0026-0363.pdf
      https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2022.869825/full
      https://www.nature.com/articles/s41556-024-01448-1

      Helpful?

  2. What are the appropriate storage conditions for Product No. A8592, Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) antibody?

    1 answer
    1. This product ships on wet ice and it is recommended to store the undiluted antibody at 0 to -20°C.  Once diluted, repeated freezing and thawing is not recommended.

      Helpful?

  3. Can product No. A8592, Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) antibody, be used for immunohistochemistry?

    1 answer
    1. The product cannot be used on paraffin embedded tissue sections, but it does work on fixed culture cells (as indicated on the Product Information Sheet) and also may work on frozen sections.

      Helpful?

  4. How is Product No. A8592, Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) antibody, supplied?

    1 answer
    1. Product No. A8592, ANTI-FLAG M2-peroxidase is supplied as a solution in 10 mM sodium phosphate, pH 7.4, containing 150 mM NaCl and 50% glycerol, plus stabilizer and preservative.  The conjugate protein concentration is approximately 1 mg/ml.

      Helpful?

  5. What is the Department of Transportation shipping information for this product?

    1 answer
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

      Helpful?

  6. I am getting a high background on my Western blots using Product No. A8592, Monoclonal Anti-FLAG® M2-Peroxidase (HRP) antibody.  What can I do to fix this?

    1 answer
    1. There is a troubleshooting guide provided at the end of the product information sheet.  Some suggestions include the following:  adjusting antibody titer, increase number of washes, increase stringency of washes by addition of 0.05% TWEEN 20 or 0.1% TRITON X-100 to the wash buffers, and/or increase the incubation and wash temperatures.

      Helpful?

  7. What concentration of Product No. A8592, Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) antibody, is recommended for my application?

    1 answer
    1. An antibody titer from 1:100 to 1:1000 is recommended for immunocytochemistry and Western blot.  Minimum antibody titer recommended for ELISA is 1:20,000.  Adjust antibody titer to maximize detection sensitivity and to minimize background.

      Helpful?

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