Mouse anti gfp

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany

The Mouse anti-GFP is a primary antibody that specifically binds to green fluorescent protein (GFP). It can be used to detect and visualize the expression of GFP-tagged proteins in various experimental systems.

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Lab products found in correlation

Rabbit anti gfp, by Thermo Fisher Scientific (21 mentions) Alexa fluor 488, by Thermo Fisher Scientific (7 mentions) Mouse anti flag, by Merck Group (6 mentions) Rabbit anti rfp, by Thermo Fisher Scientific (5 mentions) Mouse anti v5, by Thermo Fisher Scientific (4 mentions) Vectashield, by Vector Laboratories (4 mentions) Protease inhibitor cocktail, by Roche (4 mentions) Rabbit anti gaba, by Merck Group (3 mentions) Mouse anti ha, by Merck Group (3 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (3 mentions) Chicken anti gfp, by Thermo Fisher Scientific (3 mentions) Rabbit anti dsred, by Takara Bio (3 mentions) Vibratome, by Leica camera (3 mentions) Chicken anti gfp, by Abcam (3 mentions) Rabbit anti rfp, by Takara Bio (3 mentions) Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (3 mentions) 780 upright confocal microscope, by Zeiss (2 mentions) Goat anti sox2, by Santa Cruz Biotechnology (2 mentions) Rabbit anti nestin, by Merck Group (2 mentions) Chicken anti map2, by Abcam (2 mentions)

92 protocols using mouse anti gfp

Coimmunoprecipitation of Hsh49 and Hsh155

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Coimmunoprecipitations were performed using yeast strains containing TAP-tagged Hsh49 and/or GFP-tagged Hsh155 treated with or without MMS. TAP-tagged Hsh49 was captured using IgG Sepharose fast-flow beads (Sigma-Aldrich) and proceeded as described previously (Leung et al., 2016 (link)). Immunoblotting was performed with mouse anti-GFP (Thermo Fisher Scientific) and rabbit anti-TAP (Thermo Fisher Scientific).
For Western blots, whole-cell extracts were prepared by trichloroacetic acid extraction and blotted with mouse anti-GFP (Thermo Fisher Scientific) or rabbit anti-PGK1 (Abcam) essentially as described previously (Gallina et al., 2015 (link)).

Mathew V., Tam A.S., Milbury K.L., Hofmann A.K., Hughes C.S., Morin G.B., Loewen C.J, & Stirling P.C. (2017). Selective aggregation of the splicing factor Hsh155 suppresses splicing upon genotoxic stress. The Journal of Cell Biology, 216(12), 4027-4040.

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Co-IP of Yeast Protein Complexes

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Co-Immunoprecipitation was performed using yeast strains containing TAP tagged Cus1, Cdc48 and/or GFP tagged Hsh155 treated with or without MMS. TAP tagged proteins was captured using IgG sepharose fast flow beads (Sigma) and proceeded as described (33) . Immunoblotting was carried out with mouse anti-GFP (ThermoFisher) and rabbit anti-TAP (ThermoFisher).
For western blots, whole cell extracts were prepared by tricholoroacetic acid (TCA) extraction and blotted with mouse anti-GFP (ThermoFisher), rabbit anti-RFA (Agrisera) or mouse anti-PGK1 (Santa Cruz) antibodies as described (10) .

Mathew V., Kumar A., Jiang Y.K., West K., Tam A., & Stirling P.C. (2020). Cdc48 regulates intranuclear quality control sequestration of the Hsh155 splicing factor.

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Antibody Staining Protocol for Protein Detection

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Primary antibodies were as follows: anti-E-cadherin (#610181, BD Bioscience, Franklin Lakes, NJ, 1:200 dilution in immunofluorescence (IF), 1:1000 in immunobloting); anti-αTubulin (#T6074, Sigma-Aldrich, St. Louis, MO, 1/2000); mouse anti-GFP (#A-11120, Thermo Fisher, Waltham, MA, 1/100); rabbit anti-GFP (#A-11122, Thermo Fisher, 1/500); anti-active caspase-3 (#559565, BD Bioscience, 1/300); anti-β-catenin (#C7207 Sigma-Aldrich, 1/500); anti-Sephs1 (ab96542 Abcam, 1/200); and anti-mKO2 (#M168-3M, MBL, Nagoya, Japan). Secondary antibodies were as follows: AlexaFluor488-conjugated anti-mouse IgG (#A-11029, Invitrogen, Waltham, MA, 1/300) and anti-rabbit IgG (#A-11034, Invitrogen, 1/300); AlexaFluor594-conjugated anti-mouse IgG (#A-11032, Invitrogen, 1/300) and anti-rabbit IgG (#A-11037, Invitrogen, 1/300); AlexaFluor647-conjugated anti-rabbit IgG (#4414, Cell Signaling Technology, Mountain View, CA, 1/500).

Akieda Y., Ogamino S., Furuie H., Ishitani S., Akiyoshi R., Nogami J., Masuda T., Shimizu N., Ohkawa Y, & Ishitani T. (2019). Cell competition corrects noisy Wnt morphogen gradients to achieve robust patterning in the zebrafish embryo. Nature Communications, 10, 4710.

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Neuronal Morphology of NRXN1+/- hiPSC-Neurons

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Unlabeled control hiPSC-NPCs were plated onto 24-well matrigel coated plates with coverslips. Control hiPSC-NPCs were transduced with a scramble_gRNA-eGFP-puro expressing lentivirus while NRXN1^+/− hiPSC-NPCs were transduced with scramble_gRNA-tdTomato-puro expressing lentivirus. A small number of control tdTomato labeled and NRXN1^+/− eGFP-labeled hiPSC-NPCs were spiked into the unlabeled background of hiPSC-NPCs upon plating. hiPSC-NPCs were then differentiated for 6 weeks into hiPSC-neurons and immunostained using mouse anti-GFP (ThermoFisher), 1:1,000; rabbit anti-RFP (ThermoFisher) 1:1,000; with Alexa donkey anti-mouse-488 and Alexa donkey anti-rabbit-568 secondary antibodies as described above. Individual neurons were imaged at 20x on the Zeiss 780 upright confocal microscope, followed by tracing and analysis using ImageJ Simple Neurite Tracer Plugin. One-way ANOVA with Dunnett’s test was used to test the impact of genotype on neuronal morphology. As only the eGFP 3’-NRXN1^+/− were imaged on the same coverslip as tdTomato labeled controls, we assess the impact of coverslip to coverslip variation between these lines using a two-way ANOVA.

Flaherty E., Zhu S., Barretto N., Cheng E., Deans M.P., Fernando M., Schrode N., Francoeur N., Antoine A., Alganem K., Halpern M., Deikus G., Shah H., Fitzgerald M., Ladran I., Gochman P., Rapoport J., Tsankova N., Mccullumsmith R., Hoffman G.E., Sebra R., Fang G, & Brennand K.J. (2019). Neuronal impact of patient-specific aberrant NRXN1α splicing. Nature genetics, 51(12), 1679-1690.

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Immunodetection of TRPV4 and GFP

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Primary antibodies used were rabbit anti‐GFP (Cell Signaling Technology, 2555), mouse anti‐GFP (Thermo Fisher Scientific, A‐11120), rabbit anti‐TRPV4 (Cosmo Bio USA, KAL‐KM119). Secondary antibody used was HRP‐conjugated monoclonal mouse anti‐rabbit IgG, light chain specific (Jackson ImmunoResearch, 211–032‐171). Reagents used include HC067047 (Sigma‐Aldrich, SML0143), GSK2193874 (Sigma‐Aldrich, SML0942), and AlexFluor 555 Phalloidin (ThermoFisher Scientific).

Taga A., Peyton M.A., Goretzki B., Gallagher T.Q., Ritter A., Harper A., Crawford T.O., Hellmich U.A., Sumner C.J, & McCray B.A. (2022). TRPV4 mutations causing mixed neuropathy and skeletal phenotypes result in severe gain of function. Annals of Clinical and Translational Neurology, 9(3), 375-391.

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Investigating C9orf72 and TFEB Regulation

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C9orf72-Flag and C9orf72 shRNA constructs were described previously [27 (link)]. GFP-TFEB (38119, Shawn Ferguson, Yale University), pRK5-HA GST RagB (19301, David Sabatini, Whitehead Institute), pRK5-HA GST RagC (19304, David Sabatini, Whitehead Institute), myc-Raptor (1859, David Sabatini, Whitehead Institute), pRK5-HA GST RagB 99L (19303, David Sabatini, Whitehead Institute), and pRK5-HA GST RagC 75N (19305, David Sabatini, Whitehead Institute) were obtained from Addgene.
Antibodies used in the human cell studies include the following: mouse anti-Flag (Sigma), mouse anti-GFP, rabbit anti-GAPDH (ThermoFisher), mouse anti-actin (Santa Cruz), mouse anti-C9orf72 (Bio-Rad), mouse anti-C9orf72 (Proteintech), mouse anti-HA (Bethyl Lab), rabbit anti p-70S6K, rabbit anti p-p70S6K, rabbit anti-RagA, rabbit anti-RagC, rabbit anti-Raptor, rabbit anti-LAMP1, rabbit anti-DYKDDDDK, rabbit anti-pTFEB, rabbit anti-TFEB (Cell Signaling), mouse anti-TFEB (Mybiosources), and rabbit anti-GFP (Abcam).

Ji Y.J., Ugolino J., Zhang T., Lu J., Kim D, & Wang J. (2020). C9orf72/ALFA-1 controls TFEB/HLH-30-dependent metabolism through dynamic regulation of Rag GTPases. PLoS Genetics, 16(4), e1008738.

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Plasmid Sources and Antibodies for TLK1 and MK5 Studies

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Wild‐type human full‐length TLK1 mammalian expression plasmid was purchased from Addgene (Watertown, MA, USA, cat# 98378). Wild‐type full‐length human TLK1B bacterial expression plasmid was obtained from S. Kirubakaran, Discipline of Biological Engineering, Indian Institute of Technology, Gandhinagar, India [40 (link)]. Dominant negative human TLK1 kinase‐dead mammalian expression vector was generated as previously described [41 (link)]. pEGFP‐MK5‐C1 mammalian expression vector was a kind gift from O. Morten Seternes (Department of Pharmacology,1 Institute of Medical Biology, University of Tromsø, Tromsø, Norway) [42 (link)]. Human full‐length MK5 bacterial expression plasmid was purchased from Vector Builder (Chicago, IL, USA). The following primary antibodies were used in this study: rabbit anti‐TLK1 (Thermo Fisher, Waltham, MA, USA, cat# 720397), rabbit anti‐TLK1B (laboratory‐generated), rabbit anti‐MK5 (Cell Signaling Technology, CST, Danvers, MA, USA, cat# 7419S), mouse anti‐PRAK (Santa Cruz Biotechnology, SCBT, Dallas, TX, USA, sc‐46667), rabbit anti‐phospho‐MK5 S354 (Thermo Fisher, cat# PA5‐105676), mouse anti‐GFP (Thermo Fisher, cat# MA5‐15256), rabbit anti‐GAPDH (CST, cat# 2118S) and rabbit anti‐actin (Abcam, Cambridge, MA, USA, cat# ab1801).

Khalil M.I., Singh V., King J, & De Benedetti A. (2022). TLK1‐mediated MK5‐S354 phosphorylation drives prostate cancer cell motility and may signify distinct pathologies. Molecular Oncology, 16(13), 2537-2557.

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Whole-Mount Immunohistochemistry of Drosophila Brains

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For whole mount immunohistochemistry, fly brains were fixed in 4% paraformaldehyde (PFA) for 30 min, dissected, and blocked in 5% normal goat serum for 1 hr at RT. Primary and secondary antibodies were incubated at 4°C overnight. The following primary antibodies were used: rabbit anti-GFP (Molecular Probes, Eugene, OR, Cat# A-21312, RRID:AB_221478) at 1:500; mouse anti-GFP (Thermo Fisher Scientific, Waltham, MA, RRID:AB_221568) at 1:500; rabbit anti-RFP (Rockland Cat, Limerick, PA, # 600-401-379, RRID:AB_2209751) at 1:500; and mouse anti-BRP (DSHB, Iowa City, IA, Cat# nc82, RRID:AB_2314866) at 1:150. The secondary antibodies, Alexa Fluor 488 goat anti-rabbit (Thermo Fisher Scientific, Waltham, MA, Cat# A11008, RRID:AB143165), Alexa Fluor 568 goat anti-rabbit (Thermo Fisher Scientific, Waltham, MA, Cat# A11011, RRID:AB_143157), and Cy5 goat anti-mouse (Thermo Fisher Scientific, Waltham, MA, Cat# A10524, RRID:AB_2534033) were used at 1:1000. Images were obtained on a Leica SP8 confocal microscope.

Duhart J.M., Baccini V., Zhang Y., Machado D.R, & Koh K. (2020). Modulation of sleep-courtship balance by nutritional status in Drosophila. eLife, 9, e60853.

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Immunocytochemistry of hiPSC-derived Neurons

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hiPSC-NPCs, hiPSC-neurons and NGN2-neurons were fixed in 4% paraformaldehyde in PBS at room temperature for 10 min. hiPSC-NPCs were permeabilized at room temperature for 15 min in 1.0% Trition in PBS. All cells were blocked in 5% donkey serum with 0.1% Triton at room temperature for 30 min. Primary antibodies, including rabbit anti-NESTIN (Millipore), 1:500; goat anti-SOX2 (Santa Cruz), 1:500; chicken anti-MAP2 (Abcam), 1:500–1:5,000; mouse anti-SYN1 (Millipore), 1:500; mouse anti-GFP (ThermoFisher), 1:1,000; rabbit anti-RFP (ThermoFisher), 1:1,000, rabbit anti-GABA (Sigma) 1:500; were incubated overnight at 4 °C. Secondary antibodies including Alexa donkey anti-rabbit 488, 568, 647 (Invitrogen), Alexa donkey anti-mouse 488, 568, 647 (Invitrogen) and Alexa donkey anti-chicken 647 (Invitrogen) were used at 1:500–1:1,000. DAPI (0.5 μg/mL) was used to visualize nuclei. Coverslips were mounted with Vectashield and imaged using the Zeiss upright 780 confocal microscope.

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Immunofluorescence Staining of RNAi-Treated Cells

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RNAi treated cells were placed on poly-L-Lysine coated coverslips, fixed with 4% paraformaldehyde, blocked for 1 hour with standard PBS containing 10% normal goat serum (NGS) at room temperature (RT), then stained with Alexa Flouro 568 phalloidin (1:1000; #A12380, Molecular Probes, Inc., Oregon, USA) in PBS with 10% NGS for 45 min at room temperature (RT). A minimum of eight frames of fluorescence micrographs was taken containing on average 30–40 cells counted and categorized for cell shape for each treatment. For the transfected cells, the appropriate primary antibody: mouse anti-GFP (1:200; #MS-1315 Thermo Fisher Scientific, USA), rabbit anti-FLAG (1:200; #F7425 Sigma-Aldrich, Missouri, USA), mouse anti-HA (1:200; #H9658 Sigma-Aldrich, Missouri, USA) was added overnight in PBS with 10% NGS at 4°C. Cells were next incubated in PBS containing 10% NGS and secondary anti-mouse or anti-rabbit FITC (1:200) antibody. After two hours, cells were washed and stained with Alexa Flouro 568 phalloidin (1:1000; #A12380, Molecular Probes, Inc., Oregon, USA) in PBS with 10% NGS for 45 min at RT before mounting. Cells that were FITC-positive were counted and characterized.

Ojelade S.A., Acevedo S.F., Kalahasti G., Rodan A.R, & Rothenfluh A. (2015). RhoGAP18B Isoforms Act on Distinct Rho-Family GTPases and Regulate Behavioral Responses to Alcohol via Cofilin. PLoS ONE, 10(9), e0137465.

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