The primary antibodies used in the immunostaining experiments were as follows: mouse anti-GFP antibody (1:500, Roche, no. 11814460001), chicken anti-mCherry antibody (1:300, Novus Biologicals, no. NBP2-25158) and mouse anti-BRP antibody (1:100, Developmental Studies Hybridoma Bank, no. nc82). The secondary antibodies for immunostaining were anti-mouse labelled by Alexa 488 (1:1,000, ThermoFisher no. A28175) or Cy3 (1:1,000, Jackson ImmunoResearch Labs no. 115-165-146) and anti-chicken labelled by Alexa 647 (1:1,000, Jackson ImmunoResearch Labs, no. 103-605-155). The primary antibodies used in the western blots were mouse anti-GFP antibody (1:2,000, Roche, no. 11814460001), rabbit anti-mCherry antibody (1:2,000, Abcam, no. ab167453), rabbit anti-Ref2P antibody (1:500, Abcam, no. ab178440), rabbit anti-TMEM63A antibody (1:200, Novus Biologicals, no. NBP2-57359), mouse anti-GAPDH antibody (1:2,000, Proteintech, no. 60004-1-Ig) and mouse anti-tubulin antibody (1:2,000, Sigma, no. T9026). The secondary antibodies for the western blots were anti-mouse HRP antibody (1:4,000, Jackson ImmunoResearch Labs, no. 115-035-146) and anti-rabbit HRP antibody (1:4,000, Jackson ImmunoResearch Labs, no. 111-035-144).
Mouse anti gfp antibody
Manufactured by Roche
Sourced in Switzerland, Germany
The Mouse anti-GFP antibody is a laboratory reagent used to detect and quantify the presence of green fluorescent protein (GFP) in biological samples. It is a monoclonal antibody produced in mice that specifically binds to GFP, allowing for the identification and localization of GFP-tagged proteins.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Mouse anti tubulin antibody, by Merck Group (3 mentions)
X ray film, by Fujifilm (2 mentions)
Rabbit anti ha antibody, by Merck Group (2 mentions)
H6908, by Merck Group (2 mentions)
Mab1501, by Merck Group (2 mentions)
Dynabeads protein g, by Thermo Fisher Scientific (2 mentions)
Phosstop phosphate inhibitor cocktail, by Roche (2 mentions)
Horseradish peroxidase conjugated anti rabbit secondary antibody, by Fortis Life Sciences (2 mentions)
Mouse ant gfp antibody, by Roche (2 mentions)
Bis sulfosuccinimidyl suberate bs3, by Thermo Fisher Scientific (2 mentions)
Anti mouse secondary antibody, by Bio-Rad (2 mentions)
Complete lysis m, by Roche (2 mentions)
Nitrocellulose membrane, by Bio-Rad (2 mentions)
Clarity western ecl substrate, by Bio-Rad (2 mentions)
Mini protean tgx precast gel, by Bio-Rad (2 mentions)
Mouse anti ubq antibody p4d1, by Santa Cruz Biotechnology (2 mentions)
Rabbit anti mcherry antibody, by Abcam (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Lsm 700 confocal microscope, by Zeiss (2 mentions)
46 protocols using mouse anti gfp antibody
1
Immunostaining and Western Blot Antibodies
2
Viral Protein Expression and Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transient expression of two selected viral proteins fused to GFP, N, and NSs following agroinfiltration was validated by western blot. Soluble proteins from agroinfiltrated leaf tissues at 2 dpi (for localization assay) or 3 dpi (suppression of RNA silencing assay) were extracted in protein sample buffer (Laemmli, 1970 (link)) and separated on a SDS-12% polyacrylamide gel. Proteins were transferred to PVDF membrane (Millipore) and detected using mouse anti-GFP antibodies (1:1000, Roche Life Science) or mouse monoclonal anti-Flag M2 clone 2 antibody (1:2000, Sigma–Aldrich) followed by rabbit anti-mouse IgG horseradish peroxidase conjugate (1:25,000), Pierce ECL Plus chemiluminescent substrate, and exposure to x-ray film (Fujifilm). Protein sizes were estimated using pre-stained Page Ruler protein ladder (Thermo Fisher Scientific).
3
Quantifying Protein Expression in Caenorhabditis Elegans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Animals were either hand-picked, or rinsed from plates and washed in M9 buffer, and flash frozen. Prior to SDS-PAGE, thawed animals were resuspended in SDS-PAGE loading buffer, subjected to 3 min sonication in an ultrasonic bath (Ultrawave, 100 W) and heated to 95°C for 5 min. SDS-PAGE was performed using NuPAGE 4–12% gradient gels with MOPS SDS as buffer system (Invitrogen). Proteins were transferred onto Hybond-P membranes (Cytiva) by electro blotting and membranes were probed as indicated with either mouse anti-GFP antibodies (Roche 11814460001), mouse anti-mNeonGreen antibodies (Chromotek) or mouse anti-GAPDH antibodies (Invitrogen AM4300). Proteins were detected using HRP-linked anti-mouse IgG antibodies (Cell Signalling Technology) with Immobilon Forte Western HRP substrate (Merck) and visualised using an iBright FL1000 imaging system (Invitrogen). Membranes were also stained with amido black (Merck). Pre-stained broad range molecular weight marker was from New England Biolabs. Quantitation was done using Fiji (24 (link)).
4
Immunoprecipitation of GFP-Tagged Proteins
5
Antibody Dilutions for Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies were used at the following dilutions: mouse anti-GFP antibodies (1:2000) and anti-HA-Peroxidase (1:5000) were purchased from Roche. Rat anti-AGO2 monoclonal antibody (1:200), rat anti-AGO1 monoclonal antibody (1:10) and rat anti-TNRC6B antibody (1:20) were kindly provided by G. Meister. Rabbit anti-DIS3L2 (1:1000) is a gift from A. Dziembowski and rabbit anti-STARPAP (TUT1) was kindly provided by R.A. Anderson and used at 1:3300. Rabbit anti-DIS3 (1:1000), rabbit anti-TUT7 (1:500), mouse anti-tubulin (1:5000), anti-mouse, anti-rat and anti-rabbit secondary antibodies (1:10000) were purchased from Sigma-Aldrich. Rabbit anti-XRN2 (1:500), anti-EXOSC3 (1:500), anti-RRP6 (1:3000) and mouse anti-Histone H3 (1:5000) were purchased from Abcam. Rabbit anti-XRN1 (1:2000) is from Bethyl laboratories.
6
Immunostaining Protocols for Ovary and Embryo
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ovary and embryo immunostaining protocols used was previously described50 (link)51 (link). The primary antibodies used were as follows: mouse anti-GFP antibody (1:100; Roche), rabbit anti-Osk antibody (1:100; a gift from Dr. Tze-Bin Chou), rabbit anti-Tudor antibody (1:100; a gift from Dr. Akira Nakamura), rabbit anti-Krimp antibody (1:500; a gift from Dr. Toshie Kai) and rat anti-Vas antibody (1:100; Developmental Studies Hybridoma Bank). The fluorescently labelled secondary antibodies used were as follows: goat anti-mouse Alexa Fluor 488 (1:100; Invitrogen), goat anti-rabbit Alexa Fluor 647 (1:100; Invitrogen) and goat anti-rat Alexa Fluor 647 (1:100; Invitrogen).
7
Mitochondrial Localization of Apoaequorin-GFP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neurospheres were fixed with phosphate-buffered saline (PBS) containing 4% paraformaldehyde for 40 min in normal incubation conditions (i.e., at 30°C and 5% CO 2 ).
They were then immunolabeled as described previously (Dong et al. 2017 , Aulestia et al. 2018) . The correct localization of apoaequorin-GFP to the mitochondria was confirmed by the co-localisation of GFP and TOM20 signals. TOM20 is a 20 kDa translocase of the mitochondrial outer membrane (Eliyahu et al. 2010) . TOM20 was detected using the rabbit anti-TOM20 antibody (Santa Cruz Biotechnology, at a 1:100 dilution). To enhance the EGFP signal, samples were also labeled with a mouse anti-GFP antibody (Roche; at a 1:1,000 dilution). The secondary antibodies (Alexa Fluor-488 goat anti-mouse IgG and Alexa Fluor-555 goat anti rabbit IgG; Molecular Probes, Invitrogen, Eugene, OR, USA) were both used at 1:1,000 dilution. Images of immunolabeled cells were acquired with a Leica TCS SP5 II laser scanning confocal microscope mounted on a Leica DMI 6000 microscope using a Leica HCX PL APO 40/1.25 N.A. oil immersion lens. EGFP fluorescence was captured using 488 nm excitation and 510-550 nm detection; TOM20 fluorescence was captured with 561 nm excitation and 570-620 nm detection; and DAPI fluorescence was captured using 405 nm excitation and 430-480 nm detection.
They were then immunolabeled as described previously (Dong et al. 2017 , Aulestia et al. 2018) . The correct localization of apoaequorin-GFP to the mitochondria was confirmed by the co-localisation of GFP and TOM20 signals. TOM20 is a 20 kDa translocase of the mitochondrial outer membrane (Eliyahu et al. 2010) . TOM20 was detected using the rabbit anti-TOM20 antibody (Santa Cruz Biotechnology, at a 1:100 dilution). To enhance the EGFP signal, samples were also labeled with a mouse anti-GFP antibody (Roche; at a 1:1,000 dilution). The secondary antibodies (Alexa Fluor-488 goat anti-mouse IgG and Alexa Fluor-555 goat anti rabbit IgG; Molecular Probes, Invitrogen, Eugene, OR, USA) were both used at 1:1,000 dilution. Images of immunolabeled cells were acquired with a Leica TCS SP5 II laser scanning confocal microscope mounted on a Leica DMI 6000 microscope using a Leica HCX PL APO 40/1.25 N.A. oil immersion lens. EGFP fluorescence was captured using 488 nm excitation and 510-550 nm detection; TOM20 fluorescence was captured with 561 nm excitation and 570-620 nm detection; and DAPI fluorescence was captured using 405 nm excitation and 430-480 nm detection.
8
GFP-Fusion Protein Immunoprecipitation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK 293T cells in 35 mm diameter wells were transfected using Lipofectamine 2000 (Thermo Fisher Scientific). Twenty four hours posttransfection, cells were rinsed with PBS and resuspended in 400 μl NP buffer (50 mM Tris–Cl pH 7.5, 150 mM NaCl, and 1% Nonidet P40) with protease inhibitor mix (Roche Diagnostic), PMSF (0.1 mg/ml), and RNase A (10 ng/μl). After vortexing for 30 s, cell extracts were clarified by centrifugation at 20,000g for 10 min at 4 °C. The soluble extract was then incubated with 1.2 μg of a mouse anti-GFP antibody (Roche Diagnostic, Cat. Nr. 11814460001) for 1 h at 4 °C and next with 30 μl of protein G Sepharose beads (Cytiva). Beads were washed four times with washing buffer (50 mM Tris–Cl pH 7.5, 150 mM NaCl, and 0.1% Nonidet P40) before resuspension in Laemmli buffer for SDS PAGE analysis. UHMK1 was revealed with the rat monoclonal antibody 3B12 (11 ) (hybridoma supernatant at 1:20 dilution). GFP-fusion proteins were detected with a rabbit anti-GFP antibody (Santa Cruz SC8334, 1:1000). Primary antibodies were then detected with IRDye680 or IRDye800 secondary antibodies (Li-Cor). Western blots were imaged with Typhoon infrared laser (Amersham, GE Healthcare). Alternatively, gels were imaged using Pro-Q Diamond (Thermo Fisher Scientific) and imaged on the laser scanner at 532 nm. Gels were then restained with Coomassie Blue before scanning at 680 nm.
9
Western Blot Analysis of GFP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Yeast cells were washed, pelleted, and resuspended and boiled in 1 x Laemmli sample buffer to obtain extracts. After transfer, blots were developed using mouse anti-GFP antibody (Roche, 1:1000 dilution) followed by HRP-conjugated goat anti-mouse antibody (Cytiva NA931, 1:10 000 dilution) and Western Bright chemiluminescent detection reagent (advanstra). ChemiDoc MP (BioRad) was used for detection.
10
Polysome Profiling Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fractions (16 s) were collected from a polysome profiling experiment of either exponential or PBS stressed DHH1-YFP tagged cell lysate (60 OD260 cell extract per gradient). Ten microlitres of every third fraction was mixed with 2x Laemmli buffer, denatured at 95 °C for 2 min, loaded on a 10% SDS-PAGE gel, and run for 1 h at 160 V. The gel was transferred onto a nitrocellulose membrane (Amersham Biosciences) and blocked for 1 h in 1x PBS containing 0.1% Tween-20 in 5% nonfat dry milk. The membranes were incubated with mouse anti–GFP antibody (1:1000, Roche; cat. number: 11814460001) at 4 °C overnight. After washing (3 × 10 min each with 1 x PBS, 0.1 % Tween-20) horseradish peroxidase conjugated secondary antibodies were added for 1 h at room temperature (1:3000; Roche). The membranes were washed as before and results were visualized using an enhanced chemiluminescence SuperSignal West Femto Maximum Sensitivity Substrate (ThermoFisher Scientific).
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!