The largest database of trusted experimental protocols

Anti mcm2 polyclonal antibody n 19

Manufactured by Santa Cruz Biotechnology

The Anti-Mcm2 polyclonal antibody N-19 is a lab equipment product manufactured by Santa Cruz Biotechnology. It is a polyclonal antibody that specifically targets the Mcm2 protein.

Automatically generated - may contain errors

5 protocols using anti mcm2 polyclonal antibody n 19

1

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein extracts were prepared essentially as described previously [40 (link)]. Briefly, cells grown to exponential phase were incubated with YPD or SD medium containing 2 μg/ml tunicamycin, 4 mM DTT or 0.4 M sodium chloride, for the indicated times. Cells were transferred into test tubes, mixed 1:1 with boiled medium, submerged in the boiling water for 3 min, and harvested by centrifugation. Cells were then subjected to a mild alkali treatment-based protein extraction method [41 (link)]. Western blot analysis was performed using the immunoreaction enhancer solution Can Get Signal (Toyobo) according to the manufacturer's protocol. Anti-HA monoclonal antibody 16B12 (Covance), anti-Myc monoclonal antibody 9E10 (Santa Cruz), anti-GFP monoclonal antibody JL-8 (Clontech), anti-phospho-p38 MAPK monoclonal antibody 28B10 (Cell Signaling), anti-phospho-AMPKα monoclonal antibody 40H9 (Cell Signaling), anti-Hog1 polyclonal antibody y-215 (Santa Cruz), anti-Snf1 polyclonal antibody yk-16 (Santa Cruz), and anti-Mcm2 polyclonal antibody N-19 (Santa Cruz) were used. Detection was carried out by using a LAS-4000 (Fuji Film) with Immobilon Westren (Merck Millipore). Signal intensities were quantified by ImageQuant (GE Healthcare), and statistical analysis was performed with Excel (Microsoft).
+ Open protocol
+ Expand
2

Quantitative Protein Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Preparation of protein extracts and Western blot analysis were performed as described previously19 (link). Anti-GFP monoclonal antibody JL-8 (Clontech), anti-phospho-p38 MAPK monoclonal antibody D3F9 (Cell Signaling), anti-Hog1 polyclonal antibody y-215 (Santa Cruz), anti-phospho-p44/42 MAPK polyclonal antibody (Cell Signaling), anti-Mpk1 polyclonal antibody yN-19 (Santa Cruz), and anti-Mcm2 polyclonal antibody N-19 (Santa Cruz) were used. Detection was carried out by using a LAS-4000 (Fuji Film) with Immobilon Westren (Merck Millipore). Signal intensities were quantified by ImageQuant (GE Healthcare), and statistical analysis was performed with Excel (Microsoft).
+ Open protocol
+ Expand
3

Western Blot Analysis of GFP-tagged Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Preparation of protein extracts and Western blot analysis were performed as described previously [10 (link)]. WIDE RANGE Gel Preparation Buffer(4x) for PAGE (Nakalai) was used to detect degradation of GFP-tagged proteins. Anti-GFP monoclonal antibody JL-8 (Clontech), anti-GFP antibody from mouse IgG1κ (clones 7.1 and 13.1) (Roche), anti-phospho-AMPKα monoclonal antibody 40H9 (Cell Signaling), anti-Snf1 polyclonal antibody yk-16 (Santa Cruz), anti-Myc monoclonal antibody 9E10 (Santa Cruz) and anti-Mcm2 polyclonal antibody N-19 (Santa Cruz) were used. Detection was carried out by using a LAS-4000 (Fuji Film) with Immobilon Western (Merck Millipore) or the Odyssey Imaging Systems (LI-COR Biosciences). Signal intensities were quantified by the Odyssey Imaging Systems, and statistical analysis was performed with Excel (Microsoft).
+ Open protocol
+ Expand
4

Western Blot Protein Analysis Workflow

Check if the same lab product or an alternative is used in the 5 most similar protocols
Preparation of protein extracts and Western blot analysis were performed as described previously9 (link). Anti-phospho-p38 MAPK monoclonal antibody D3F9 (Cell Signaling), anti-Hog1 polyclonal antibody y-215 (Santa Cruz), anti-phospho-p44/42 MAPK polyclonal antibody (Cell Signaling), anti-Mpk1 polyclonal antibody yN-19 (Santa Cruz), anti-Myc monoclonal antibody 9E10 (Santa Cruz) and anti-Mcm2 polyclonal antibody N-19 (Santa Cruz) were used. Detection was carried out by using a LAS-4000 (Fuji Film) with Immobilon Western (Merck Millipore) or the Odyssey Imaging Systems (LI-COR Biosciences). Signal intensities were quantified by the Odyssey Imaging Systems, and statistical analysis was performed with Excel (Microsoft).
+ Open protocol
+ Expand
5

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cells collected from indicated times were then treated with sodium hydroxide for protein extraction, as described previously [44 (link)]. Protein samples were loaded on to an 8% or 10% SDS-PAGE gel for protein electrophoresis and then transferred to a PDVF membrane (Millipore) for Western blot analysis. Anti-FLAG polyclonal antibody M2 (Sigma), anti-Mcm2 polyclonal antibody N-19 (Santa Cruz), anti-Mcm4 polyclonal antibody yC-19 (Santa Cruz), anti-Mcm7 polyclonal antibody yN-19 (Santa Cruz), and anti-Elm1 polyclonal antibody y-640 (Santa Cruz) were used to detect 3Flag-Lrg1, Mcm2, Mcm4, Mcm7, and Elm1, respectively. The monoclonal anti-Pgk1 antibody 22C5D8 (Invitrogen) was used to detect Pgk1, as the loading control, since Pgk1 is reported to be a very stable protein based on its half-life [45 (link)]. Detection was carried out by using a LAS-4000 (Fuji Film) with Immobilon Western (Merck Millipore). Signal intensities were quantified by means of Image Quant (GE Healthcare).
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!