Neutrophils (3 × 106) from blood were isolated by percoll density gradients as previously described [3 (link)] and were lysed with a boiling Laemmli buffer. Samples were loaded onto a 15% SDS-PAGE gel. Proteins were then transferred onto nitrocellulose membranes using Bio-Rad’s Trans-Blot Turbo, which were blocked using 5% nonfat dry milk in 1X TBS-T buffer for 1 h at room temperature. The membranes were incubated overnight at 4 °C under mild agitation in 5% nonfat dry milk in 1X TBS-T buffer containing the following primary antibodies: mouse anti-Caspase-1 (p20) (mAb (Bally-1); Adipogen; AG-20B-0048-C100; 1:1000), mouse anti-Caspase-4 (MBL cat. M029-3; 1:1000), rabbit anti-GSDMD (Abcam, cat. ab215203; 1:1000), rabbit anti-α-actin (Sigma-Aldrich, cat. A2066; 1:5000), mouse anti-α-actin (Cell Signaling, cat. 3700; 1:1000), rabbit anti-RIG-I (D14G6) (Cell Signaling, cat. 3743S; 1:1000). Membranes were washed in 1x TBS-T and incubated with appropriate secondary HRP-conjugated antibodies diluted in 5% nonfat dry milk in 1X TBS-T buffer. Protein detection was done using an ECL™ Prime Western Blotting System (GE Healthcare) and an Amersham Imager 600 (GE Healthcare).
Mouse anti α actin
Manufactured by Cell Signaling Technology
Mouse anti-α-actin is a primary antibody that specifically recognizes the α-actin isoform. It can be used to detect and quantify the expression of α-actin in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using mouse anti α actin
1
Neutrophil Protein Expression Analysis
2
Neutrophil Protein Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neutrophils (3 x 10 6 ) from blood were isolated by percoll density gradients as previously described (3) and were lysed with a boiling Laemmli buffer. Samples were loaded onto a 15% SDS-PAGE gel. Proteins were then transferred onto nitrocellulose membranes using Bio-Rad's Trans-Blot Turbo, which were blocked using 5% nonfat dry milk in 1X TBS-T buffer for 1 h at room temperature. The membranes were incubated overnight at 4°C under mild agitation in 5% nonfat dry milk in 1X TBS-T buffer containing the following primary antibodies: mouse anti-Caspase-1 (p20) (mAb (Bally-1); Adipogen; AG-20B-0048-C100; 1:1000), mouse anti-Caspase-4 (MBL cat. M029-3; 1:1000), rabbit anti-GSDMD (Abcam, cat. ab215203; 1:1000), rabbit anti-α-actin (Sigma-Aldrich, cat. A2066; 1:5000), mouse anti-α-actin (Cell Signaling, cat. 3700;
. 1:1000), rabbit anti-RIG-I (D14G6) (Cell Signaling, cat. 3743S; 1:1000). Membranes were washed in 1x TBS-T and incubated with appropriate secondary HRP-conjugated antibodies diluted in 5% nonfat dry milk in 1X TBS-T buffer. Protein detection was done using an ECL™ Prime Western Blotting System (GE Healthcare) and an Amersham Imager 600 (GE Healthcare).
. 1:1000), rabbit anti-RIG-I (D14G6) (Cell Signaling, cat. 3743S; 1:1000). Membranes were washed in 1x TBS-T and incubated with appropriate secondary HRP-conjugated antibodies diluted in 5% nonfat dry milk in 1X TBS-T buffer. Protein detection was done using an ECL™ Prime Western Blotting System (GE Healthcare) and an Amersham Imager 600 (GE Healthcare).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!