Anti-KLF4 抗体 (ab106629)

Run in running buffer at 130V for 2hours(or until the dye front is close to the bottom);

Transfer the proteins to the membrane at 250mA in transfer buffer for 2hours.

After transfer, wash membrane in 1xTBST. 

Incubate the membrane with blocking buffer overnight at 4°C with gentle agitation. 

Dilute ab106629 to recommended dilution in 10mL of blocking buffer. Incubate membrane with ab106629 overnight at 4^oC. Wash 3 times x 10 minutes in washing buffer with gentle agitation.

Dilute 1μL anti-rabbit IgG-HRP conjugated secondary in 10mL of blocking buffer to make a 1:10,000 dilution (can vary from 1:2,000 to 1:10,000). Incubate membrane for 1 hour at room temperature. Wash 3 times x 10 minutes in washing buffer with gentle agitation. 

Running Buffer: 25mM Tris, 192 mM Glycine, 0.1% SDS.

Transfer Buffer: 20mM Tris, 150mM Glycine, 20% Methanol, 0.038% SDS.

Blocking Buffer: 5% non-fat dry milk, TBS.

Wash Buffer (TBST): 125 mM NaCl, 25mM Tris pH 8.0, 0.1% Tween-20. 

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling KLF4 with ab106629 at 5 µg/mL. Antigen Retrieval: Use Citrate buffer antigen retrieval using a microwave. Endogenous peroxide blocking with 3% H₂O₂. Blocking with 5% BSA at room temperature for 30 minutes. ab106629 incubation at 4°C overnight. Secondary antibody incubation at room temperature for 30 minutes. Washing 3 times for 5 minutes using phosphate buffered saline. Detection using DAB.