P. falciparum parasites were released from erythrocytes by treatment with 0.15% saponin in PBS. Murine erythrocytes infected with the pv5-tag-GFPPV or exp2-mCherry P. berghei lines (62 (link)) were purified on a Nycodenz gradient and lysed hypotonically for 1 h on ice in 10 mM Tris⋅HCl, pH 7.5. P. berghei lysates were centrifuged for 50 min at 100,000 × g. Membrane pellets were resuspended in 0.1 M Na2CO3 in PBS or in 1% Triton X-100 in PBS, respectively, and centrifuged for 50 min at 100,000 × g. Proteins were separated on SDS-polyacrylamide and transferred to nitrocellulose membranes. Western blotting was performed using rat anti-mCherry (1:5,000; ChromoTek), chicken anti-GFP (1:5,000; Abcam), rat anti-HA (1:1,000; Sigma Aldrich), rat anti-PfBiP (1:1,000) (63 (link)), and rabbit anti-human hemoglobin α-primary antibodies (1:1,000; Abcam) followed by chemiluminescence detection with horseradish peroxidase-coupled secondary antibodies (1:10,000; Sigma Aldrich, or 1:5,000; Jackson ImmunoResearch).
Rat anti mcherry
Manufactured by Proteintech
The Rat anti-mCherry is a primary antibody that specifically binds to the mCherry protein. mCherry is a red fluorescent protein commonly used as a reporter or tag in various biological applications. The Rat anti-mCherry antibody can be used to detect and visualize mCherry-tagged proteins in cell-based assays, immunohistochemistry, or other relevant experiments.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti rec 8, by Novus Biologicals (2 mentions)
Rabbit anti him 8, by Novus Biologicals (2 mentions)
Horseradish peroxidase coupled secondary antibody, by Merck Group (1 mentions)
Rat anti ha, by Merck Group (1 mentions)
Chicken anti gfp, by Abcam (1 mentions)
Rabbit anti rad 51, by Novus Biologicals (1 mentions)
Mouse anti ha, by Cell Signaling Technology (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Roche (1 mentions)
Tank blot device, by Bio-Rad (1 mentions)
Mouse anti gfp, by Roche (1 mentions)
Image lab software 5, by Bio-Rad (1 mentions)
Rabbit anti gfp, by Thermo Fisher Scientific (1 mentions)
Chemidoc xrs imaging system, by Bio-Rad (1 mentions)
Rat anti ha, by Roche (1 mentions)
5 protocols using rat anti mcherry
1
Purification and Western Blot Analysis of Plasmodium Proteins
2
Validated Immunostaining Antibodies for C. elegans
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The following primary antibodies were used at the indicated dilutions: rabbit anti-GFP-488-conjugated (1:200) (Invitrogen, A21311), goat anti-GFP-FITC-conjugated (1:200) (Abcam, ab6662), rat anti-mCherry (1:1000) (Chromotek, 5F8), rabbit anti-COH-3/4 (1:400)21 (link), mouse anti-REC-8 (1:100) (Novus Biologicals, 29470002), guinea pig anti-SYP-1 (1:400)26 (link), chicken anti-SYP-1 (1:300)47 (link), guinea pig anti-HTP-3 (1:800)32 (link), rabbit anti-HIM-3 (1:400)33 (link), rabbit anti-HTP-1/2 (1:400)47 (link), rabbit anti-PLK-2 (1:500)65 (link), guinea pig anti-SUN-1 pS12 (1:1000)19 (link), rabbit anti-RAD-51 (1:10000) (Novus Biologicals, 29480002), mouse anti-HA (1:200) (Cell Signalling, 23675), rabbit anti-DSB-2 (1:1000)9 (link), rabbit anti-HIM-8 (1:500) (Novus Biologicals, 41980002). Phospho-specific antibodies against HIM-8 pT64 (1:400) were produced by injecting rabbits with the synthetic peptide DTPRFSpTPIVPNVC (GenScript). Polyclonal anti-HIM-8 pT64 antibodies were affinity purified by binding to a column containing the phospho-peptide and the specificity of the antibodies was validated by absence of staining in germlines of chk-2 mutant worms.
3
Western Blot Analysis of NrC4a CRISPR Constructs
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Western blot was performed on N. benthamiana leaves transiently expressing slnrc4aCRISPR constructs19 . One hundred milligrams of plant tissue were ground with liquid nitrogen, and three volumes of extraction buffer (50 mM of Tris‐HCl, pH 7.5, 2 mM of MgCl2, 150 mM of NaCl, 140 mM of β‐mercaptoethanol, 2 mM of phenylmethylsulfonyl fluoride (PMSF), and one complete protease inhibitor tablet, without EDTA [Roche, Germany] per 50 ml) were added. Samples were centrifuged, and supernatant cytosolic fraction was discarded (or collected if required). Pellets were ground using two volumes of EB with 1% Triton X‐100 and incubated in a rotating wheel at 4 °C for 20 min before centrifugation. Supernatant samples (TSM) were collected and boiled after adding sample buffer (8% sodium dodecyl sulfate, 40% glycerol, 200 mM of Tris‐Cl, pH 6.8, 388 mM of dithiothreitol, and 0.1 mg ml−1 of bromophenol blue dye). Samples were run in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, blotted onto nitrocellulose membranes, and incubated rat anti‐mCherry (Chromotek).
4
Immunostaining of Meiotic Proteins in Caenorhabditis elegans
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The following primary antibodies and dilutions were used: goat anti-GFP-488-conjugated (1:200) (Roche), rat anti-mCherry (1:1000) (5F8, ChromoTek), rabbit anti-COH-3/4 (1:400) (Crawley et al., 2016 (link)), mouse anti-REC-8 (1:100) (Novus Biologicals), guinea pig anti-HCP-6 (1:400) (Chan et al., 2004 (link)), rabbit anti-HIM-8 (1:500) (Novus Biologicals), and rabbit anti-RAD-51 (Das et al., 2022 (link)).
5
Western Blot Analysis of Parasite Proteins
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Protein samples were resolved by SDS-PAGE and transferred to nitrocellulose membranes (Protran) in a tankblot device (Bio-Rad) using transfer buffer (0.192 M Glycine, 0.1% SDS, 25 mM Tris) with 20% methanol. Blocking of membranes and dilutions of antibodies were done in PBS containing 5% skim milk. Washing steps were done with PBS. Primary antibodies were applied in the following dilutions: mouse anti-GFP (Roche) 1:1000; rabbit anti-GFP (Thermo) 1:2000; rat anti-mCherry, 1:1000 (Chromotek); rabbit anti-SERA5, 1:2000 (newly raised); rabbit anti-REX3, 1:2000 (newly raised); mouse anti-SBP1N, 1:2500 (newly raised); rabbit anti-aldolase, 1:4000 (newly raised); mouse anti-HSP101, 1:1000 [7 (link)]; rabbit anti-DHFR (Abcam), 1:1000; rat anti-HA (Roche) 1:4000. Horseradish peroxidase-conjugated secondary antibodies used were goat anti-rat (Dianova) and goat anti-mouse (Dianova) and diluted 1:3000 as well as donkey anti-rabbit (Dianova) 1:2500 and applied after three washes. Immunoreactions were detected by enhanced chemiluminiscence (Bio Rad/ Thermo) and detected on CEA RP NEW x-ray films (Agfa). For quantification of Western blot signals, band intensities were measured with a Chemi Doc XRS imaging system (Bio-Rad) and densitometry analyses were done with Image Lab Software 5.2 (Bio-Rad). Data are representative of three independent experiments.
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