Product # | Size | Price |
---|---|---|
13089S | 100 µl | $ 276 |
REACTIVITY | H M R |
SENSITIVITY | Endogenous |
MW (kDa) | 43-48 |
Source/Isotype | Mouse IgG2b |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Immunofluorescence (Immunocytochemistry) | 1:400 - 1:800 |
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 262
This protocol is intended for immunoprecipitation of native proteins utilizing Protein G agarose beads for subsequent analysis by western immunoblot or kinase activity.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
10X Cell Lysis Buffer: (#9803) To prepare 10 ml of 1X cell lysis buffer, add 1 ml cell lysis buffer to 9 ml dH2O, mix.
NOTE: Add 1 mM PMSF (#8553) immediately prior to use.
Proceed to one of the following specific set of steps.
NOTE: To minimize masking caused by denatured IgG heavy chains (~50 kDa), we recommend using Mouse Anti-Rabbit IgG (Light-Chain Specific) (L57A3) mAb (#3677) or Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127). To minimize masking caused by denatured IgG light chains (~25 kDa), we recommend using Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127).
posted October 2016
revised April 2018
Protocol Id: 1244
Achieve higher quality immunofluorescent images using the efficient and cost-effective, pre-made reagents in our #12727 Immunofluorescence Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Recommended Fluorochrome-conjugated Anti-Mouse secondary antibodies:
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 148
RUNX3/AML2 (D9K6L) Mouse mAb recognizes endogenous levels of total RUNX3 protein.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with recombinant protein surrounding Gly217 of human Runx3 protein.
Runt-related transcription factor 3 (RUNX3, AML2), a member of the Runt family of transcription factors, plays an important role in the suppression of gastric epithelium cell proliferation (1), dorsal root ganglia neurogenesis (2), and T cell differentiation (3,4). RUNX3 is also involved in caspase-3-dependent apoptosis (5). Protein complexes containing RUNX3 and various transcription factors, such as Smads or β-catenin/TCF4, have tumor suppressor activity and regulate downstream target gene transcription (6,7). While typically localized to the nucleus, RUNX3 can be tyrosine phosphorylated and located in the cytoplasm of many cancer cells. This mislocalization of RUNX3 abolishes its tumor suppressor function and contributes to tumorigenesis (8). Research studies indicate that gene silencing or protein mislocalization inactivates RUNX3 in more than 80% of gastric cancers and other cancer types (1,9,10).
Explore pathways + proteins related to this product.
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