LS4-2G12 (1:5000 IHC; 1:2000 IF) and 9C10 (1:10,000) are mouse monoclonal antibodies raised to pSer129 epitope of αSyn and total synuclein (residues 2–21) respectively (Dhillon et al., 2017 (link)). Other antibodies used include: rabbit anti-p62 (1:2000, ProteinTech), mouse anti-ubiquitin (1:1000, EMD Millipore), rabbit Iba-1 (1:1000, Wako), mouse Cd68 (1:200, Invitrogen), rabbit anti-GFAP (1:1000, Dako), and rabbit anti-cd11b (1:1000, AbCam).
Mouse anti ubiquitin
Manufactured by Merck Group
Sourced in United States
Mouse anti-ubiquitin is a primary antibody reagent used in research applications. It binds to ubiquitin, a small regulatory protein involved in various cellular processes. This antibody can be utilized in techniques such as Western blotting, immunoprecipitation, and immunohistochemistry to detect and study ubiquitin-related mechanisms.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti pgk1, by OriGene (2 mentions)
Mouse anti gfp, by Roche (2 mentions)
Gfp trap coupled agarose beads, by Proteintech (2 mentions)
Rabbit anti p62, by Proteintech (1 mentions)
Rabbit iba1, by Fujifilm (1 mentions)
Rabbit anti cd11b, by Abcam (1 mentions)
Rabbit anti gfap, by Agilent Technologies (1 mentions)
Protein g magnetic beads, by Merck Group (1 mentions)
Complete tablets edta free, by Roche (1 mentions)
Phosstop tablet, by Roche (1 mentions)
Immobilon fl pvdf membrane, by Merck Group (1 mentions)
Rabbit anti phospho stat3 tyr 705 antibody, by Cell Signaling Technology (1 mentions)
Revert total protein stain kit, by LI COR (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Rabbit anti gfp, by Thermo Fisher Scientific (1 mentions)
Multi gauge, by Fujifilm (1 mentions)
Las 3000 luminescent image analyzer, by Fujifilm (1 mentions)
Protease inhibitor mix, by Roche (1 mentions)
Dylight 488 conjugated anti mouse igg, by Abcam (1 mentions)
Rabbit anti mcherry, by Takara Bio (1 mentions)
8 protocols using mouse anti ubiquitin
1
Monoclonal Antibodies for Alpha-Synuclein
2
Immunoprecipitation and Western Blot of SERCA1 Ubiquitination
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293 cells were washed with PBS and subsequently lysed in RIPA solubilization buffer (50 mM TRIS-HCl pH 7.4, 150 mM NaCl, 1% NP40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate) supplemented with 2 mM N-ethyl-maleimide (NEM) (Sigma) and protease inhibitors. 450 μg of total-cell lysate (input) was subjected to immunoprecipitation. Samples were incubated overnight, tumbling at 4 °C with the polyclonal antibodies specific to SERCA1 (Cell Signaling, Danvers, MA, USA). The following day, protein G-magnetic beads (Millipore, Darmstadt, Germany) were added and incubated for 1 h at 4 °C. Beads were recovered, extensively washed with RIPA buffer containing N-ethyl-maleimide (NEM) and aspirated to dryness. SERCA1 was eluted by heating beads at 70 °C in sample-loading buffer with added beta-mercaptoethanol for subsequent detection by Western blotting. The blots were blocked with Tris-buffered saline, Tween 20 (TBS-T) with 5% milk powder and probed with mouse anti-ubiquitin (Merck Millipore, Burlington, MA, USA) and then incubated with secondary HRP-conjugated antibodies (Vector Laboratories, Newark, CA USA). Bands were detected as described above. Images were acquired by the Alliance mini HD9 Imaging System.
3
Western Blot Analysis of Ubiquitin Chains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We employed the following antibodies: rabbit anti-UbK63 chains (Millipore), mouse anti-ubiquitin (Chemicon), mouse anti-γH2AX (clone JBW301, Upstate), rabbit anti-γH2AX (Epitomics), rabbit anti-53BP1 (Alexis). Following the indicated treatments whole cell lysates were prepared using lysis buffer containing: 50 mM Tris-HCl [pH 7.4], 150 mM NaCl, 1% NP-40, 0.5% Sodium Deoxycholate, 1 mM EDTA, 0.1% SDS, supplemented with protease- (EDTA-free Complete tablets, Roche) and phosphatase inhibitors (PhosSTOP tablets, Roche). Supernatants were boiled in Laemmli buffer and proteins were resolved by SDS-PAGE. Proteins were transferred onto PVDF membranes and blocked for 1 hour in 5% skim milk in TBS, 0.05% Tween-20 (TBS-T). Membranes were probed overnight at 4°C with antibodies directed against K63 chains (1∶1000) or ubiquitin (1∶500). Bound antibodies were visualized using HRP-linked secondary antibodies (anti-rabbit (GE Healthcare) and anti-mouse (GE Healthcare)) and ECL luminescence (Pierce).
4
Detailed Western Blot Procedure for Ubiquitin and pSTAT3
5
Western Blot Antibody Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blots were performed according to standard procedures. The following primary antibodies were used: rabbit anti-Mask (1:1,000),14 (link) mouse anti-mCherry antibody (1:1000, Novus Biologicals, clone 1C51, NBP1–96752), mouse anti-ubiquitin (1:1000, Millipore, clone FK2, 04–263), rabbit anti-K48-Ub (1:1000, Cell Signaling Technology, clone D9D5, mAb #8081), rabbit anti-K63-Ub (1:500, Millipore, clone Apu3, 05–1308), rabbit anti-GFP (1:1000, Life Technologies, A11122), rabbit anti-Ref(2)P (1:10,000), mouse anti-ACTA/α-actin (1: 5000, JLA20) and mouse anti-TUBB/β-tubulin (1:1,000, E7) were obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at the University of Iowa, Department of Biology, Iowa City, IA. HRP-conjugated anti-rabbit or HRP-conjugated anti-mouse secondary antibodies were used at 1:10000. Data were collected using Luminescent Image Analyzer LAS-3000 (FUJIFILM, Valhalla, NY) and quantified using Multi Gauge (FUJIFILM, Valhalla, NY).
6
Purification of GFP-Tagged Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gim3-TAP yeast cells transformed with a control empty vector (BPM 42), PGPD-GFP (BPM 779), PGPD-Guk1-GFP (BPM 780), or PGPD-Guk1-7-GFP (BPM 781) were grown to log phase and then lysed with glass beads and native lysis buffer (20 mM HEPES, pH 7.5, 0.5% NP-40, 200 mM NaCl, 1X protease inhibitor mix, 1 mM 1,10 phenanthroline, 1 mM EDTA, 10 mM iodoacetamide). To pulldown GFP-tagged proteins, lysates were incubated for 2 hours at 4°C with 20 μL GFP-Trap coupled agarose beads (Chromotek). Beads were washed three times in lysis buffer before samples were eluted with 3X SDS sample buffer. Nitrocellulose membranes were probed with mouse anti-GFP (Roche, 1:2,500), rabbit anti-Pgk1 (Acris Antibodies, 1:10,000), rabbit anti-TAP (Fisher, 1:2,500), and mouse anti-ubiquitin (Millipore, 1:2,500) primary antibodies.
7
Guk1-7 Protein Interaction Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells expressing ectopic Guk1-7-GFP, Guk1-GFP, or a control empty vector (pRS313) were grown to log phase and then lysed with glass beads in native lysis buffer (20 mM HEPES, pH 7.5, 0.5% NP-40, 200 mM NaCl, 1X protease inhibitor mix (Roche), 1 mM 1,10 phenanthroline, 1 mM EDTA, 10 mM iodoacetamide). GFP-tagged Guk1-7 was pulled down with GFP-Trap coupled agarose beads (Chromotek; 10 µL per 3 mg of lysate) for 2 hours at 4°C. Beads were washed three times in lysis buffer before samples were eluted with 3X SDS buffer. Equal volumes of samples were resolved by SDS-PAGE. Membranes were immunoblotted with mouse anti-GFP (Roche, 1:2,500), rabbit anti-Pgk1 (Acris Antibodies, 1:10,000), and mouse anti-ubiquitin (Millipore, 1:2,500) primary antibodies and secondary antibodies (Mandel Scientific, 1:10,000).
8
Immunocytochemistry of Larval Muscles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Larval muscles were dissected in ice-cold phosphate-buffered saline (PBS; 137 mM NaCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 2.7 mM KCl, pH 7.4) and fixed in 4% paraformaldehyde for 30 min. For the detection of autofluorescence from proteins tagged with fluorescent molecules, the fixed tissues were washed with PBS, and mounted for imaging. For immunocytochemistry, the fixed tissues were stained following standard procedures. The primary antibodies used were: mouse anti-ubiquitin at 1:1000 (Millipore, Clone FK2, 04–263), rabbit anti-mCherry at 1:1000 (Clontech, 632496), Alexa Fluor 488-conjugated rabbit anti-GFP at 1:1000 (Life Technologies, A21311), rat anti-HA at 1:1000 (Roche, 11867431001). The following secondary antibodies were used: Cy3-conjugated goat anti-rabbit IgG at 1:1000 (Jackson ImmunoResearch, 111–165–144), Dylight 488 conjugated anti-mouse IgG at 1:1000 (Abcam, ab150113).
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!