Antibodies used in the study were as follows: anti-Flag (F3165/F7425, Sigma), anti-Myc (9B11/71D10, Cell Signaling Technology (CST)), anti-HA (16B12, Biolegend and H6908, Sigma), anti-TRAF2 (sc-136999, Santa Cruz), anti-Rab5 (sc-46692, Santa Cruz), anti-Rab7 (sc-376362, Santa Cruz), anti-Sprouty 2 (sc-100862, Santa Cruz/ab85670, Abcam), anti-EGFR (sc-373746, Santa Cruz), anti-p-Tyr (9411, CST), anti-p-Ser/Thr (ab9344, Abcam and 05-368, Sigma), anti-Ser (sc-81514, Santa Cruz), anti-Thr (sc-5267, Santa Cruz), anti-Ubiquitin (sc-8017, Santa Cruz), anti-β-Actin (AC026, Abclonal), anti-GFP (AE011, Abclonal), anti-glutathione S-transferase (GST) (2622 S, CST), anti-V5 (R960-25, Thermo Fisher and 30801ES10, Yeasen), anti-Mouse/Rabbit IgG-peroxidase secondary antibody (A0545/A9044, Sigma), anti-Mouse IgG (light chain specific)-peroxidase secondary antibody (115-005-174, Jackson), and anti-DYRK1A polyclonal antibody has been previously described [30 (link)].
Anti gfp
Manufactured by ABclonal
Sourced in China
The Anti-GFP is a laboratory tool that is used to detect and quantify the presence of green fluorescent protein (GFP) in biological samples. It is a specific antibody that binds to GFP, allowing researchers to identify and analyze GFP-tagged proteins or cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti flag, by Merck Group (7 mentions)
Anti actin, by ABclonal (3 mentions)
Protease inhibitor cocktail, by Roche (3 mentions)
Anti myc, by ABclonal (2 mentions)
Anti gst, by ABclonal (2 mentions)
Anti his, by ABclonal (2 mentions)
Complete protease inhibitor cocktail, by Roche (2 mentions)
Anti flag, by ABclonal (2 mentions)
Ab85670, by Abcam (1 mentions)
Ab9344, by Abcam (1 mentions)
Anti egfr, by Santa Cruz Biotechnology (1 mentions)
H6908, by Merck Group (1 mentions)
Anti rab7, by Santa Cruz Biotechnology (1 mentions)
Anti rab5, by Santa Cruz Biotechnology (1 mentions)
Anti traf2, by Santa Cruz Biotechnology (1 mentions)
Anti sprouty 2, by Santa Cruz Biotechnology (1 mentions)
Anti ubiquitin, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by ABclonal (1 mentions)
Anti ha, by BioLegend (1 mentions)
Protein a magnetic bead, by Merck Group (1 mentions)
22 protocols using anti gfp
1
Antibody Panel for Protein Analysis
2
ChIP-qPCR Protocol for OsMYB8 in Rice
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For ChIP assays, rice florets of the ZH11 and transgenic pOsMYB8::OsMYB8-GFP plants were harvested and cross-linked in a fixation buffer with 1% (v/v) formaldehyde under vacuum for 20 min. Glycine was added to terminate the cross-linking reaction. The prepared chromatin complexes were sonicated into 200–500 bp fragments and then precleared with protein A magnetic beads (Merck Millipore, USA)64 (link). For immunoprecipitations, Anti-GFP (Nanobody) Magnetic beads (ABclonal, China) were added into samples and incubated overnight at 4 °C. DNA was precipitated with magnetic beads. For the real-time-qPCR reaction, the precipitated DNA was recovered and dissolved in water as the template. The enrichment was standardized to the input DNA to obtain the fold enrichment. An unrelated DNA sequence from the rice OsActin1 gene was used as an internal control. All relevant primers used in the ChIP assay are listed in Supplementary Data 11 .
3
Protein Extraction and Western Blot Analysis
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Two hundred milligrams of plant tissue were collected and placed into a 2 ml grinding tube (Jingxin). After grinding plant tissues at low temperature, 1 ml of protein extraction buffer (Beyotime) was added, after which the contents were shaken at 4 °C for 30 min, followed by centrifugation at 13,000g and 4 °C for 15 min. The centrifuged supernatant was homogenized with a bicinchoninic acid kit (CWBIO), mixed with an equal volume of 2× SDS loading buffer and denatured at 100 °C for 8 min. The proteins were resolved using 10 to 20% SDS‒PAGE gels (Genescript). The Western blot–specific antibodies used included anti-GFP (ABclonal), anti-HA (CST), anti-Actin (ABclonal), anti-SWP1 (ABclonal), and HRP goat anti-rabbit immunoglobulin G (H + L) (ABclonal).
4
Western Blot Protein Detection Protocol
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Proteins were separated on SDS‐PAGE gels (10%), before being transferred to Immune‐Blot PVDF membranes (Roche). Subsequently, the membranes were incubated in blocking buffer for 2 h at 4°C. After blocking, the membranes were incubated with anti‐His (#AE003, ABclonal), anti‐GFP (#AE011, ABclonal), anti‐GST (#AE006, ABclonal), anti‐MBP (#AE016, ABclonal), anti‐myc (#AE009, ABclonal), anti‐actin (#AC009, ABclonal) antibodies for 2 h at 4°C. The membranes were subsequently washed with blocking buffer for three times, before being incubated with the secondary antibody HRP goat anti‐rabbit IgG (H + L) antibody (#AS014, ABclonal), or HRP goat anti‐mouse IgG (H + L) antibody (#AS003, ABclonal). Finally, the membranes were washed with TBST and they were developed using the eECL western blot kit (CWBIO, Beijing, China).
5
Western Blot Analysis of Yeast Lysates
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Yeast extracts were prepared using TCA lysis method as described above. Either equal amounts or a serial dilution of the lysates was prepared from various samples before resolving them in SDS-PAGE and transferring onto a polyvinylidene difluoride membrane. Following incubation with primary rabbit or mouse antibody and corresponding HRP-conjugated secondary antibody, protein signals were detected by chemiluminescence using Pierce ECL Plus Western Blotting Substrate (Thermo Scientific) and autoradiography. The following antibodies were used with their source and catalog numbers indicated within parentheses: anti-Flag M2 (F3165; Sigma), anti-Pgk1 (459250; Invitrogen), anti-GFP (AE011, Abclonal), anti-UBE2A (A7744; Abclonal), anti-UBE2B (A6315, Abclonal), anti-V5 (R690, Invitrogen), anti-H2B (39237; Active Motif), anti-H3 (ab1791; Abcam), anti-H3K4me1 (39297; Active Motif), anti-H3K4me2 (399141; Active Motif), anti-H3K4me3 (39159; Active Motif), anti-ubiquitin antibody (ab139467; Abcam); monoubiquitinylated and polyubiquitinylated conjugates monoclonal antibody (FK2) (HRP conjugate) (BML-PW0150; Enzo Life Sciences), and anti-Rad6 (DZ33919; Boster Bio). Please note that the anti-UBE2B antibody recognizes both UBE2A and UBE2B (Fig. S2 ).
6
CoIP of ULT1 and TCP14/15/DA1 Proteins
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CoIP experiments were performed as described previously (Dong et al., 2020 (link)). In brief, 14-day-old 35S::ULT1-FLAG and 35S::ULT1-FLAG 35S::TCP14/TCP15/DA1-GFP/YFP seedlings were collected, and their total proteins were extracted with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 5 mM EDTA [pH 8.0], 0.1% Triton X-100, 0.2% NP-40, 1 nM freshly added PMSF, complete protease inhibitor cocktail [Roche], and 10 μM MG132). After centrifugation, protein A beads (Invitrogen) were incubated with the supernatant for 4 h, and anti-GFP antibody was then added and co-incubated overnight. The beads were washed three times with wash buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 20% glycerol, 0.1% Triton X-100, 1 mM EDTA [pH 8.0], complete protease inhibitor cocktail [Roche], and 10 μM MG132). The immunoprecipitates were separated by 10% SDS–PAGE and detected by immunoblot analysis with anti-FLAG (Sigma) and anti-GFP (Abclonal) antibodies.
7
Western Blotting with Custom Antibodies
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Western blotting was performed according to standard protocols. The primary antibodies used in this study were obtained as follows: anti-GFP (ABclonal Technology; AE012), anti-β-tubulin (Yeasen Tech; 30303ES50), anti-NONHSAT130014+unORF+2+peptide9, and anti-NONHSAT077882+1+orf4 were customized and raised by GeneScript Biotech Corporation.
8
Immunoblot Analysis of Bacterial Effector Proteins
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Total soluble proteins were extracted from N. benthamiana leaves expressing RipAW, RipE1, INF1, NahG or RipAW’s derivatives. Proteins were separated by SDS-PAGE and transferred onto PVDF membranes (Thermo Fisher) which were then blocked using 5% defatted milk in TBST buffer (1× TBS containing 0.05% Tween 20) for 60 min at room temperature. The membranes were incubated with anti-FLAG (1:8000, Sigma), anti-HA (1:10000, Abmart) or anti-GFP (1:8000, ABclonal) at room temperature for 1 h and then with a horseradish peroxidase-conjugated secondary antibody (1:10000, Sigma) for another 45 min. Signals were detected and photographed using the SuperSignal West Pico Chemiluminescent substrate (Thermo Scientific) and the Bio-Rad ChemiDoc Touch Imaging System.
9
Western Blotting of HEK293T Cells
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For Western blotting, 24-well plate HEK293T cells were lysed by RIPA. The antibodies used included anti-Cas9 (Genscript (A01935, clone 4A1), 1:500), anti-GAPDH (Santa cruz (sc47724, clone 0411), 1:1000), and anti-GFP (ABclonal (AE012), 1:2000). Images were captured with Amersham Imager 600. Uncropped blots for the presented results are provided in the Source Data file.
10
Western Blot Protein Analysis
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Total proteins were extracted with RIPA lysis buffer (Solarbio), separated by 10% SDS-PAGE, and then transferred onto polyvinylidene difluoride (PVDF) membranes and blocked using 5% nonfat milk powder (Santa Cruz Biotechnology) for 2 h. The membrane was then incubated with primary antibody, followed by HRP-labeled secondary antibody (1:1000 dilution; Santa Cruz). Protein expression was detected using a chemiluminescence detection kit (Invitrogen). The utilized antibodies were the following: anti-GFP (Abclonal AE012), anti-FLAG (Sigma-Aldrich F3165), anti-Ago2 (Abcam 186733), and anti-H3 (Active Motif sc-69970).
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