Log In Sign up
The largest database of trusted experimental protocols

Mouse anti pgk1

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in Japan

Mouse anti-Pgk1 is an antibody that binds to the Pgk1 (Phosphoglycerate kinase 1) protein. Pgk1 is an essential enzyme involved in the glycolytic pathway. The mouse anti-Pgk1 antibody can be used to detect and quantify the Pgk1 protein in various experimental applications.

Automatically generated - may contain errors

Lab products found in correlation

Odyssey infrared imager, by LI COR (2 mentions) Mouse anti gfp, by Roche (2 mentions) Rabbit anti tap, by Thermo Fisher Scientific (2 mentions) A9044, by Merck Group (1 mentions) Anti gfp, by GenScript (1 mentions) Anti rfp, by GenScript (1 mentions) Anti dpm1, by Thermo Fisher Scientific (1 mentions) Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Glass bead, by Merck Group (1 mentions) Rabbit anti clb2, by Santa Cruz Biotechnology (1 mentions) Mouse anti actin, by Thermo Fisher Scientific (1 mentions) Mouse anti gfp, by Merck Group (1 mentions) Rabbit anti gfp, by Abcam (1 mentions) Rabbit anti actin, by Abcam (1 mentions) Mouse anti flag m2, by Merck Group (1 mentions) Mouse anti myc 9e10, by Santa Cruz Biotechnology (1 mentions) Mouse anti ha, by Roche (1 mentions) Mouse anti flag, by Merck Group (1 mentions) Mouse anti c myc, by Santa Cruz Biotechnology (1 mentions)

10 protocols using mouse anti pgk1

1

Antibody Immunodetection Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following antibodies were used: mouse anti-mini-AID (1:1000; M214-7; Medical & Biological Laboratories (MBL), Aichi, Japan), mouse anti-Pgk1 (1:500, 459250, Thermo Fisher Scientific, Waltham, MA, United States), anti-mouse IgG (whole molecule) (1:5000; A9044, Sigma-Aldrich, St. Louis, MO, United States).
Nakamura T.S., Suda Y., Muneshige K., Fujieda Y., Okumura Y., Inoue I., Tanaka T., Takahashi T., Nakanishi H., Gao X.D., Okada Y., Neiman A.M, & Tachikawa H. (2021). Suppression of Vps13 adaptor protein mutants reveals a central role for PI4P in regulating prospore membrane extension. PLoS Genetics, 17(8), e1009727.
+ Open protocol
+ Expand
2

Yeast Protein Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
For protein analysis, two OD 600 units of yeast cells were harvested and frozen prior to extraction. Total proteins were extracted as described previously (Li et al, 2016) . Equal volumes of total lysates were analyzed using SDS-PAGE and transferred to a polyvinylidene difluoride (PVDF) membrane (Merck Millipore). Rabbit anti-BMV 1a antiserum (1:10,000 dilution, a gift from Dr. Paul Ahlquist at the University of Wisconsin-Madison), mouse anti-BMV 2a pol (1:3000 dilution, a gift from Dr. Paul Ahlquist at the University of Wisconsin-Madison), mouse anti-Pgk1 (1:10,000 dilution; Thermo Fisher Scientific), anti-GFP (1:5000 dilution), and anti-RFP (1:3000 dilution, GenScript), anti-Dpm1 (1:3,000 dilution, Thermo Fisher Scientific) was used as the primary antibody. Horseradish peroxidase conjugated anti-rabbit or anti-mouse antibody (1:10,000 dilution, Thermo Fisher Scientific) together with the Supersignal West Femto maximum sensitivity substrate (Thermo Fisher Scientific) was used to detect target proteins. Band intensities were quantified using Adobe Photoshop.
Sathanantham P., Zhao W., He G., Murray A., Fenech E., Díaz A.P., Schuldiner M., & Wang X. (2021). A conserved viral amphipathic helix governs the replication site-specific membrane association.
+ Open protocol
+ Expand
3

Yeast Whole-Cell Extract Preparation

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Yeast whole-cell extracts were prepared as described previously64 (link). In brief, cells at OD600 of 0.5 in log-phase were collected and washed immediately with 20% trichloroacetic acid (TCA). The cell pellet was suspended in 250 μl of 20% TCA, mixed with an equal volume of glass beads (Sigma, 425–600 μm) and vortexed for 15 min at 4 °C. glass beads were washed twice with 250 μl of 5% TCA to increase the protein yield. The resulting extract was spun at 13,000 r.p.m. for 5 min at 4 °C. The pellet was dissolved in 100 μl of 2 × SDS-loading buffer (100 mM Tris-HCl (pH 6.8), 4% SDS, 20% glycerol, 0.1% bromophenol blue, 50 mM DTT), neutralized by adding 20 μl of 1 M Tris base, boiled for 5 min and clarified by centrifugation. Total cell extracts were subjected to immunoblot analysis. Proteins were detected using the following primary antibodies: mouse anti-Pgk1 (1:10,000; Invitrogen, 459250), rabbit anti-Clb2 (1:400; Santa Cruz Biotechnology, y-180), rabbit anti-Cof1 (1:2,000), mouse anti-Actin (1:1,000, Thermo Fisher scientific, mAbGEa) and rabbit anti-Sac6 (1:2,000). Blots were subsequently scanned using Odyssey Infrared Imager (LI-COR Biosciences). Full scans are provided in Supplementary Fig. 12.
Miao Y., Han X., Zheng L., Xie Y., Mu Y., Yates JR I.I.I, & Drubin D.G. (2016). Fimbrin phosphorylation by metaphase Cdk1 regulates actin cable dynamics in budding yeast. Nature Communications, 7, 11265.
+ Open protocol
+ Expand
4

Western Blot Analysis of Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Proteins separated by SDS-PAGE were transferred to PVDF membranes using the TRANS-BLOT semidry system from BIO-RAD or the wet blotting system in transfer buffer (25 mM Tris, 192 mM glycine, 0.1% SDS, 20% methanol, pH 8.3). The membranes were blocked using a Li-Cor blocking buffer with an overnight incubation at room temperature. Incubation with primary antibodies was performed at 4 °C for at least 1 h. The membranes were incubated in Li-Cor Odyssey secondary antibodies at room temperature for at least 1 h and then revealed in an Odyssey scanner. The primary antibodies used were mouse anti-PGK1 (Invitrogen; dilution 1:10,000), rabbit anti-TAP-TAG (Thermo Scientific; dilution 1:10,000), mouse anti-GFP (Sigma; dilution 1:2000), and rabbit anti-Asc1 (dilution 1:5000) (86 (link)). The secondary antibodies that were used were anti-mouse 680 LT Li-Cor (dilution 1:10,000) and anti-rabbit 800 CW Li-Cor (dilution 1:10,000).
Barros G.C., Requião R.D., Carneiro R.L., Masuda C.A., Moreira M.H., Rossetto S., Domitrovic T, & Palhano F.L. (2021). Rqc1 and other yeast proteins containing highly positively charged sequences are not targets of the RQC complex. The Journal of Biological Chemistry, 296, 100586.
+ Open protocol
+ Expand
5

Protein Extraction and Immunoblot Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in 1 ml 0.2 M NaOH/ 0.2% β-mercaptoethanol and 100 μl trichloroacetic acid (TCA) was added. Precipitated proteins were harvested via centrifugation at 12,000 rpm and resuspended in 100 μl 2X SDS sample buffer. 20 μl of 1 M Tris base (pH ~11) was then added, samples were heated at 75°C and analyzed via immunoblot (Peng and Weisman, 2008 (link)). For immunoblot analyses, mouse anti-GFP (1:1,000; Roche), rabbit anti-GFP (1:1,000; abcam), rabbit anti-TAP (1:1,000; Thermo Scientific), mouse anti-Pgk1 (1:10,000; Invitrogen), sheep anti-Vac17 (1:1,000) and rabbit anti-phosphoThr240 (1:2,500) antibodies were used.
Yau R.G., Peng Y., Valiathan R.R., Birkeland S.R., Wilson T.E, & Weisman L.S. (2014). Release from myosin V via regulated recruitment of an E3 Ub ligase controls organelle localization. Developmental cell, 28(5), 520-533.
+ Open protocol
+ Expand
6

Analyzing Snf1 Phosphorylation in Yeast

Check if the same lab product or an alternative is used in the 5 most similar protocols
To prevent activation of SNF1 by processing of the cells (Wilson et al., 1996 (link)), cells were killed before centrifugation by addition of 1ml 100% TCA to 5ml of cells. Cells were vortexed with glass beads, pelleted, and resuspended in non-fluorescent sample buffer (Licor 928-40004). Protein concentration was determined by the Coomassie elution with SDS method (Marbach et al., 2001 (link)), except that destaining was done with water. Protein extracts were run on 10% (12% or 15% for Mms21-3HA) TGS gels (Biorad) and transferred to PVDF membranes. Phostag (Waco) was added at 0.15ml/10 mix when required. Membranes were probed with mouse anti-Myc (9E10, Santa Cruz), rabbit anti-S-tag (Abcam), rabbit anti-phospho-RxxS (Cell Signaling), mouse anti-HA (Roche or Santa Cruz), mouse anti-FLAG (M2, Sigma), with all antibodies diluted 1000-fold in blocking buffer (Rockland MB-070). Snf1 phosphorylated on T210 (P-Snf1) was detected using rabbit anti-phospho T172 AMPKα (Cell Signaling). Loading controls were detected with rabbit anti-actin (Epitomics, 500-fold diluted) or mouse anti-Pgk1 (Invitrogen, 10000-fold diluted) antibodies. Secondary antibodies were anti-mouse 680LT and anti-rabbit 800CW (LiCOR) or anti-mouse Dylight 488 and anti-rabbit Dylight 549 (Epitomics). Blots were visualized with a LiCOR odyssey or a Biorad imager. Quantifications were performed using NIH ImageJ.
Simpson-Lavy K.J., Bronstein A., Kupiec M, & Johnston M. (2015). Cross-talk between carbon metabolism and the DNA damage response in S. cerevisiae. Cell reports, 12(11), 1865-1875.
+ Open protocol
+ Expand
7

Monitoring AID*-tagged Protein Degradation

Check if the same lab product or an alternative is used in the 5 most similar protocols
To determine the timing of IAA-dependent Cin8-AID* and Boi1-AID* degradation, DDY5584, DDY5658, and DDY5655 precultures were diluted to OD600 0.1 in imaging medium (Synthetic Complete medium lacking tryptophan and with 2% glucose) and grown to OD600 0.5, at which point 250 µM IAA was added. An equal amount of cells was harvested for every time point during the 120-min time course. Cells were immediately precipitated with TCA as previously described (Foiani et al., 1994 (link)), and 0.5 OD600s were loaded into each lane. The cell extracts were subjected to SDS-PAGE and immunoblot analysis using mouse anti-FLAG (Sigma-Aldrich) and mouse anti-Pgk1 (Invitrogen) antibodies.
Ibarlucea-Benitez I., Ferro L.S., Drubin D.G, & Barnes G. (2018). Kinesins relocalize the chromosomal passenger complex to the midzone for spindle disassembly. The Journal of Cell Biology, 217(5), 1687-1700.
+ Open protocol
+ Expand
8

Controlled DNA Double-Strand Break Formation

Check if the same lab product or an alternative is used in the 5 most similar protocols
The integrated GAL10-HO cassette W303 yeast strain (MATa leu2::GAL-HO-LEU2 hmlΔ hmrΔ RAD5) was used to control DSB formation at the MAT locus (Herskowitz & Jensen, 1991) (gift from L. Symington). The mre11 gene of the parental HO yeast strain was replaced with 13x myc-tagged wild type (gift from D. Durocher) or mutated mre11 genes following established protocols [62 ]. The kanMX gene (cloned from the pFA6a-kanMX6 vector; Addgene plasmid # 39296 [63 (link)]) was juxtaposed to the Mre11 gene for the antibiotic selectivity, and the mre11Δ knock-out strain was prepared by replacing the mre11 open reading frame with kanMX gene. Integrated gene sequences were verified by PCR and western blot. Protein extracts for western blot analysis were prepared by TCA precipitation. Antibodies: mouse anti-c-myc (Santa Cruz, 9E10), mouse anti-PGK1 (Invitrogen, 22C5D8), goat anti-mouse-AP (Invitrogen, 31320). PGK1 was used as the loading control. Immunodetection was conducted using an alkaline phosphatase/NBT/BCIP system (Pierce).
Rahman S., Beikzadeh M., Canny M.D., Kaur N, & Latham M.P. (2020). Mutation of Conserved Mre11 Residues Alter Protein Dynamics to Separate Nuclease Functions. Journal of molecular biology, 432(10), 3289-3308.
+ Open protocol
+ Expand
9

Protein Extraction and Immunoblot Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were harvested and lysed in 1 ml 0.2 M NaOH and 0.2% β-mercaptoethanol. Proteins were precipitated via the addition of 100 µl TCA and centrifuged at 12,000 rpm. Proteins were resuspended in 120 µl 2× SDS sample buffer followed by the addition of 30 µl of 1 M Tris base, pH ∼11. Samples were heated at 75°C and analyzed via immunoblot (Peng and Weisman, 2008 (link)). For immunoblot analyses, mouse anti-GFP (1:1,000; Roche), rabbit anti-TAP (1:1,000; Thermo Fisher Scientific), mouse anti-Pgk1 (1:10,000; Invitrogen), sheep anti-Vac17 (1:1,000; custom made; 21st Century Biochemicals), rabbit anti–phospho-Thr240 (1:2,500; custom made; 21st Century Biochemicals), and rabbit anti–phospho-Ser222 (1:2,500; custom made; 21st Century Biochemicals) antibodies were used (see the Antibody preparation section).
Yau R.G., Wong S, & Weisman L.S. (2017). Spatial regulation of organelle release from myosin V transport by p21-activated kinases. The Journal of Cell Biology, 216(6), 1557-1566.
+ Open protocol
+ Expand
10

Immunoblot Analysis of Yeast Cell Extracts

Check if the same lab product or an alternative is used in the 5 most similar protocols
Yeast total cell extracts were prepared from log-phase cells as previously described (Foiani et al., 1994 (link)). Total cell extracts were subjected to immunoblot analysis. Proteins were detected using the following primary antibodies: mouse anti-Hrr25 (Abmart, Berkeley Heights, NJ), rabbit anti-GFP (Molecular Probes, Eugene, OR), mouse anti-V5 (Invitrogen, Carlsbad, CA), mouse anti-Pgk1 (Invitrogen). Blots were subsequently scanned using an Odyssey Infrared Imager (LI-COR Biosciences, Lincoln, NE).
Peng Y., Moritz M., Han X., Giddings T.H., Lyon A., Kollman J., Winey M., Yates J I.I.I., Agard D.A., Drubin D.G, & Barnes G. (2015). Interaction of CK1δ with γTuSC ensures proper microtubule assembly and spindle positioning. Molecular Biology of the Cell, 26(13), 2505-2518.
+ 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!