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Purified anti-MAD2 Antibody

Pricing & Availability
Clone
Poly23A13 (See other available formats)
Regulatory Status
RUO
Other Names
Mitotic spindle assembly checkpoint protein MAD2A, mitotic arrest deficient 2-like protein 1, MAD2L1, HsMAD2
Isotype
Rabbit Polyclonal IgG
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Product Citations
publications
Poly23A13_PURE_MAD2_Antibody_1_011224
Whole cell extracts (15 µg total protein) from the indicated cell lines or recombinant proteins (cell-free) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.1 µg/mL of Purified anti-MAD2 (clone Poly23A13) for 2 hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP Donkey anti-rabbit IgG (Cat. No. 406401). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control for the cell lysates (Daudi and HeLa cells) at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • Poly23A13_PURE_MAD2_Antibody_1_011224
    Whole cell extracts (15 µg total protein) from the indicated cell lines or recombinant proteins (cell-free) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.1 µg/mL of Purified anti-MAD2 (clone Poly23A13) for 2 hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP Donkey anti-rabbit IgG (Cat. No. 406401). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control for the cell lysates (Daudi and HeLa cells) at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • Poly23A13_PURE_MAD2_Antibody_2_011224
    HeLa cells were fixed with 4% PFA Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained without (Panels A and B) or with (Panels C and D) 5 µg/mL of Purified anti-MAD2 (clone Poly23A13), followed by incubation with Alexa Fluor® 594 Donkey anti-rabbit IgG (Cat. No. 406418) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (Panels A and C). The images were captured on a Revvity Operetta CLS™ High Content Analysis System with a 40X objective. Scale bar: 50 µm
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627651 50 µL £226
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627652 200 µL £449
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Description

MAD2 (mitotic arrest deficient 2) is a spindle checkpoint protein that regulates the progression of mitosis. First identified in yeast, there exist two orthologues in humans, MAD2L1 and MAD2L2, whose expression patterns are varied and often overlap. During mitosis MAD2 is recruited to unattached kinetochores and sequesters CDC20, preventing the activation of APC (anaphase promoting complex) and blocking progression from metaphase to anaphase. Thus, MAD2 acts as a check against aneuploidy by preventing the improper segregation of sister chromatids. This antibody is specific to the MAD2L1 homolog, not MAD2L2.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Polyclonal
Host Species
Rabbit
Immunogen
Recombinant full-length human MAD2L1
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.05 - 0.25 µg/mL. For immunocytochemistry, a concentration range of 5.0 - 10 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This antibody (clone Poly23A13) is not suitable for immunohistochemistry (IHC).

This antibody (clone Poly23A13) is cross-reactive with mouse MAD2/MAD2L1 in western blot (WB). However, several prominent non-specific bands are observed around 42 kD when detecting mouse samples.

By western blot (WB) this antibody recognizes only MAD2/MAD2L1, not its homolog MAD2L2.

For immunocytochemistry (ICC), fixation with 4% PFA followed by permeabilization with 0.5% Triton-X is recommended. Fixation and permeabilization with 100% ice-cold methanol alone, or fixation with 4% PFA followed by permeabilization with 100% ice-cold methanol is not recommended due to mis-localization of signal.

RRID
AB_3097526 (BioLegend Cat. No. 627651)
AB_3097526 (BioLegend Cat. No. 627652)

Antigen Details

Structure
MAD2L1 is a 205 amino-acid protein with a predicted molecular weight of 23.5 kD.
Distribution

Primarily nuclear, kinetochores

Function
Mitosis, Spindle checkpoint
Interaction
Mad2l2, cdc20, APC
Biology Area
Cell Biology, Cell Motility/Cytoskeleton/Structure, Neuroscience, Neuroscience Cell Markers
Antigen References
  1. Krishnan R, et al. 1998. Genomics. 49:475-8.
  2. Díaz-Martínez LA, et al. 2009. Cell Cycle. 8:2733-40.
  3. Hardwick KG. 2005. Curr Biol. 15:R122-4.
Gene ID
4085 View all products for this Gene ID
UniProt
View information about MAD2 on UniProt.org
Go To Top Version: 1    Revision Date: 01/12/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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