REACTIVITY | SENSITIVITY | MW (kDa) | Isotype |
---|---|---|---|
H M R Mk | Endogenous | 95-140 | Rabbit IgG |
Western blot analysis of extracts from various cell lines using Sec31A (D1G7I) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Learn more about how we get our imagesConfocal immunofluorescent analysis of MCF7 (left) and HCT-15 (right) cells using Sec31A (D1G7I) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Learn more about how we get our imagesFor western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or 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 (#13953, 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 November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised June 2016
Protocol Id: 263
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-Rabbit 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: 24
Application | Dilutions |
---|---|
Western Blotting | 1:1000 |
Immunofluorescence (Immunocytochemistry) | 1:400 |
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.
Sec31A (D1G7I) Rabbit mAb recognizes endogenous levels of total Sec31A protein.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser86 of human Sec31A protein.
The coat protein complex II (COPII) is composed of five cytosolic proteins and includes the Sec23/24 complex, the Sec13/31 complex, and Sar1. The COPII coat is located at the ER/Golgi interface and is involved in transport of newly synthesized proteins from the ER to the Golgi apparatus (1). COPII formation is initiated through the binding of the activated G protein, Sar1, to the Sec23/24 complex to form a pre-budding complex that directly binds target molecules (1-3). This pre-budding complex further recruits Sec13/31 to form mature COPII coat (4,5). The Sec31 subunit of COPII coat interacts with Sec13 at the ER exit and is required for both vesicle formation and ER-Golgi transport. Two isoforms of human Sec31 have been identified, Sec31A and Sec31B, which share a sequence homology of 47.3% (6-8). Sec31A is ubiquitously expressed in tissues and organs, whereas Sec31B is enriched in brain and testis (7,8). In classical Hodgkin lymphoma, a novel fusion of Jak2 with Sec31A renders Jak2 constitutively active and subject to Jak2 inhibitor effects (9).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. DRAQ5 is a registered trademark of Biostatus Limited. Tween is a registered trademark of ICI Americas, Inc.
Explore pathways related to this product.
Product # | Size | Price |
---|---|---|
13466S | 100 µl (10 western blots) | $ 255.0 |