The custom rabbit polyclonal antiserum against PARP-1 used for Western blotting and immunofluorescent staining assays was generated by using an antigen comprising the amino-terminal half of PARP-1 (Kim et al., 2004 (link)) (now available from Active Motif; cat. no. 39559). The custom recombinant antibody-like anti-ADP-ribose binding reagent was generated and purified in-house (Gibson et al., 2017 (link)) (now available from EMD Millipore; cat. no. MABE1031). The other antibodies used are as follows: DDX21 rabbit polyclonal antibody (Proteintech, 10528–1-AP), NOP58 rabbit polyclonal antibody (Invitrogen, PA5–5432), FLAG mouse monoclonal antibody (Sigma-Aldrich, F3165), RPS3 rabbit polyclonal antibody (Bethyl, A303–841A), RPL10 rabbit polyclonal antibody (BioRad, VPA00362), RPL26 rabbit polyclonal antibody (Sigma, HPA030449), puromycin mouse monoclonal antibody (Millipore, MABE343), β-actin rabbit polyclonal antibody (Cell Signaling, #4967), β-tubulin rabbit polyclonal antibody (Abcam, ab6046), SNRP70 rabbit polyclonal antibody (Abcam, ab51266), H2A.X (Ser139) mouse monoclonal antibody (Sigma, 05–636), and rabbit IgG (ThermoFisher Scientific, 10500C).
β actin rabbit polyclonal antibody
Manufactured by Cell Signaling Technology
The β-actin rabbit polyclonal antibody is a laboratory reagent used to detect the presence and quantity of the β-actin protein in biological samples. β-actin is a widely expressed cytoskeletal protein that plays a fundamental role in cell structure and function.
Automatically generated - may contain errors
2 protocols using β actin rabbit polyclonal antibody
1
Characterization of PARP-1 and ADP-ribose Antibodies
2
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was carried out as previously described [35] (link). Briefly, equal amounts of protein samples were run on 10% gel (casting system from Bio-Rad) with 1x Tris/Glycine/SDS buffer. Gels were transferred to PVDF membrane using wet transfer system (Bio-Rad) overnight at 16 V in Tris/Glycine/SDS buffer containing 20% methanol then blocked for one hour in 1% bovine serum albumin (BSA) in TBS buffer containing 0.05% Tween-20 (TBS-T). The membranes were incubated at RT for 2 h with 1:2000 Lon protease rabbit polyclonal antibody (gift from Luke Szweda), 1:2000 Hsp60 mouse monoclonal antibody (Enzo ADI-SPA-806), 1:2000 mouse monoclonal ClpP protease antibody (Sigma WH0008192M1), or 1:2000 β-Actin rabbit polyclonal antibody (Cell Signaling 4967 S). The blots were washed 3 times with TBS-T and then incubated with 1:10,000 HRP-conjugated secondary antibodies (Vector Laboratories) for 2 h. The blots were washed 3 more times with TBS-T, detected using ECL, and imaged using a G: BOX imaging system (Syngene). The image was then quantified using GeneTools software (Syngene).
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!