The proteins were isolated from liver macrophages and cultured RAW264.7 cells by lysing them in RIPA buffer containing 1% PMSF (Beyotime). The protein concentration was determined using a BCA protein kit (Beyotime). Based on the protein concentration, equal amounts of protein were separated by SDS/PAGE and transferred onto a PVDF membrane (Millipore Corp.) followed by blocking. The following primary antibodies were used in this study: anti‐STING (19851‐1‐AP; Proteintech), anti‐BAX (ab32503; Abcam), anti‐BAK (#578; CST), anti‐Bcl2 (ab182858; Abcam), anti‐p65 (#8242S; CST), anti‐p‐p65 (#3033S: CST), anti‐TBK1 (ab40676; Abcam), anti‐p‐TBK1 (#5483S; CST), anti‐caspase3 (ab184787; Abcam), anti‐IRF3 (#11904S; CST), anti‐p‐IRF3 (#37829S; CST), and anti‐β‐actin (7D2C10; Proteintech). The protein bands were visuzed using an enhanced chemiluminescence detection system (Bio‐Rad) and quantified after normzation to internal control β‐actin using the ImageJ software (NIH).
Anti β actin 7d2c10
Manufactured by Proteintech
Anti-β-actin (7D2C10) is a mouse monoclonal antibody that recognizes β-actin, a structural protein found in all eukaryotic cells. The antibody is suitable for a variety of applications, including Western blotting, immunofluorescence, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Ab40676, by Abcam (1 mentions)
Ab184787, by Abcam (1 mentions)
Ab182858, by Abcam (1 mentions)
Ab32503, by Abcam (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
Bca protein kit, by Beyotime (1 mentions)
Enhanced chemiluminescence detection system, by Bio-Rad (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Trans blot turbo, by Bio-Rad (1 mentions)
2 protocols using anti β actin 7d2c10
1
Western Blot Analysis of Macrophage Proteins
2
Immunoblotting with Flag-tagged Proteins in HEK293T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293T cells (ATCC)
were maintained in DMEM supplemented with glutamine, penicillin, streptomycin,
and fetal bovine serum (Seradigm). FlagDNAJB8H31Q plasmid has been reported previously.31 (link)FlagGAPDH in the pCMV3 vector was purchased from Sino
Biological. Immunoblots were transferred to nitrocellulose from SDS-PAGE
gels using a Bio-Rad Trans-Blot Turbo, stained with ponceau S to image
total protein, blocked with 5% dried milk/TBST (10 mM Tris pH 7.4,
150 mM NaCl, 0.1% Tween), washed with TBST, incubated with primary
antibody in 5% BSA/TBS with 0.1% NaN3, washed with TBST,
incubated with a near-IR-conjugated secondary antibody (Li-COR) in
5% dried milk/TBST, washed with TBST, washed with TBS, and imaged
on a Li-COR Fc. Mouse monoclonal M2 anti-Flag was purchased from Sigma-Aldrich.
Rabbit polyclonal anti-HSPA1A and mouse monoclonal anti-β-actin
(7D2C10) were purchased from Proteintech. Mouse monoclonal anticarboxymethyllysine
(#318003) was purchased from R&D Systems.
were maintained in DMEM supplemented with glutamine, penicillin, streptomycin,
and fetal bovine serum (Seradigm). FlagDNAJB8H31Q plasmid has been reported previously.31 (link)FlagGAPDH in the pCMV3 vector was purchased from Sino
Biological. Immunoblots were transferred to nitrocellulose from SDS-PAGE
gels using a Bio-Rad Trans-Blot Turbo, stained with ponceau S to image
total protein, blocked with 5% dried milk/TBST (10 mM Tris pH 7.4,
150 mM NaCl, 0.1% Tween), washed with TBST, incubated with primary
antibody in 5% BSA/TBS with 0.1% NaN3, washed with TBST,
incubated with a near-IR-conjugated secondary antibody (Li-COR) in
5% dried milk/TBST, washed with TBST, washed with TBS, and imaged
on a Li-COR Fc. Mouse monoclonal M2 anti-Flag was purchased from Sigma-Aldrich.
Rabbit polyclonal anti-HSPA1A and mouse monoclonal anti-β-actin
(7D2C10) were purchased from Proteintech. Mouse monoclonal anticarboxymethyllysine
(#318003) was purchased from R&D Systems.
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!