SNAI1 antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Storeasconcentrated solution. Centrifuge briefly prior to opening vial.For short-termstorage(1-2 weeks), storeat 4ºC.For long-termstorage,aliquotand storeat -20ºC or below. Avoid multiplefreeze-thaw cycles.	-1:500-1:3000

Upstream tips
-Storeasconcentrated solution. Centrifuge briefly prior to opening vial.For short-termstorage(1-2 weeks), storeat 4ºC.For long-termstorage,aliquotand storeat -20ºC or below. Avoid multiplefreeze-thaw cycles.
Protocol tips
-1:500-1:3000

Publication protocol

Cultured cells were placed on ice, washed in ice-cold PBS and lysed using RIPA lysis buffer (cat. 20-188, Millipore, MA) containing Protease Inhibitor Cocktail Set I (cat. 539131, Millipore, MA). Cells were then scraped and incubated for 20 minutes at 4 ˚C. The lysate was spun down at 14,000 g for 20 minutes at 4 °C. The supernatant was removed for protein content measurement. Tissue extracts were prepared from frozen tumor tissues. Briefly, the tissue was cut into smaller pieces, and the total protein was extracted by TOTAL PROTEIN EXTRACTION KIT (cat. 2140, Millipore, MA). The protein content of cell lysates and tissue extracts were measured using Bradford reagent (BIO-RAD, Hercules, CA). A measure of 40 μg of total cell extract was separated on 10% SDS-polyacrylamide gel (30% Acrylamide/Bis Solution, 29:1, cat. 1610157, BIO-RAD, Hercules, CA) and transferred to Poly Screen polyvinylidene difluoride (PVDF) membranes (cat. NEF1002001PK, Perkin-Elmer life sciences, Boston, MA). The blot was blocked in 4% milk in TBST (10 mM Tris-HCl, 150 mM NaCl, and 0.05% Tween 20) for 1 hour, and then incubated with goat anti-HSV-1 Thymidine Kinase antibody (1:200, cat. sc-28038, Santa Cruz, CA), rabbit anti-ALDH1A1 antibody (1:1000, cat. ab9883, Abcam, Cambridge, UK), mouse anti-CD133 antibody (1:1000, Miltenyi Biotec GmbH, Bergisch Gladbach, Gemany), rabbit anti-snail1 (1:500, cat. GTX125918, GeneTex, USA), anti-Akt/PKB antibody (cat. 05-591, Millipore, MA) or anti-phospho-Akt antibody (cat. 05-1003, Millipore, MA) as primary antibody for 4 ˚C overnight. After repeated wash in TBST, the membrane was incubated with secondary anti-goat, anti-rabbit or anti-mouse IgG antibodies conjugated to horseradish peroxidase (1:5000, Abcam, Cambridge, UK) for 1 hour at room temperature. Immunoreactive bands were detected using LumiFast Chemiluminescence Detection Kit (T-Pro Biotechnology, cat. JT96-K001S, Taiwan, ROC). Image capture was performed on an ImageQuant™ LAS 4000 (GE Healthcare Bio-Sciences Corp., USA).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Western blotting SNAI1 using SNAI1 antibody from GeneTex.

Manufacturer protocol

Download the product protocol from GeneTex for SNAI1 antibody below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting SNAI1 using SNAI1 antibody from GeneTex. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.