- 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
- Ave. Rating
- Submit a Review
- Product Citations
- publications
![Poly23A13_PURE_MAD2_Antibody_1_011224 Poly23A13_PURE_MAD2_Antibody_1_011224](https://d1spbj2x7qk4bg.cloudfront.net/Admin/Public/GetImage.ashx?Image=/Files/Images/media_assets/products/product_images/Poly23A13_PURE_MAD2_Antibody_1_011224.png&Width=240&Height=300&altFmImage_path=&Compression=90&Crop=5)
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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 -
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
Cat # | Size | Price | Quantity Check Availability | Save | ||
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627651 | 50 µL | $330 | ||||
627652 | 200 µL | $655 |
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 DetailsProduct 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
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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
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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
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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
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- Krishnan R, et al. 1998. Genomics. 49:475-8.
- Díaz-Martínez LA, et al. 2009. Cell Cycle. 8:2733-40.
- 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
Related Pages & Pathways
Pages
Related FAQs
Other Formats
View All MAD2 Reagents Request Custom ConjugationDescription | Clone | Applications |
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Purified anti-MAD2 | Poly23A13 | WB,ICC |
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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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