Anti rfp

Manufactured by Rockland Immunochemicals

Sourced in United States

Anti-RFP is a laboratory reagent used to detect and quantify the presence of red fluorescent protein (RFP) in biological samples. It functions as a specific antibody that binds to RFP, enabling researchers to measure and analyze RFP expression levels in their experiments.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (5 mentions) Anti gfp, by Abcam (4 mentions) Anti gfp, by Aves Labs (3 mentions) Alexa fluor 647 goat anti rabbit igg h l, by Thermo Fisher Scientific (2 mentions) Anti pcna, by Santa Cruz Biotechnology (2 mentions) Alexa fluor 488 goat anti mouse igg h l, by Thermo Fisher Scientific (2 mentions) Lsm 700 microscope, by Zeiss (2 mentions) Anti gfp, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Anti iba1, by Fujifilm (2 mentions) Sample buffer, by Bio-Rad (2 mentions) Anti alix, by Proteintech (2 mentions) Ecl select western blotting detection reagent, by GE Healthcare (2 mentions) Chemidoc mp imaging system, by Bio-Rad (2 mentions) Avidin biotin blocking kit, by Vector Laboratories (2 mentions) Anti ttf1, by Abcam (2 mentions) Avidin biotin complex kit, by Vector Laboratories (2 mentions) Anti tp63, by Cell Signaling Technology (2 mentions) Anti uchl1, by Merck Group (2 mentions) Ipfl00010, by Merck Group (2 mentions)

46 protocols using anti rfp

Immunohistochemical Analysis of Mouse Brain

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Adult mice were transcardially perfused with paraformaldehyde (4%) to fixate tissue and brains were extracted. Gelatin-embedded slices of 40 μm thick were blocked in 3% H₂O₂ in PBS, rinsed, and for antigen retrieval incubated in sodium citrate (10 mM) at 80°C for 20 min. The slices were again rinsed and placed in blocking solution (10% normal horse serum and 0.5% Triton X-100 in PBS) for 1h at room temperature and incubated overnight at 4°C in blocking solution with AffiniPure Fab Fragment (donkey anti mouse IgG (H+L), 1:200, Cat# 715-007-003, Jackson ImmunoResearch). The next day, slices were incubated for 48–72h with primary antibodies (anti-CAMKII alpha (6G9 clone), 1:100, NB100-1983, Novus Biologics; anti-RFP, 1:5000, 600-401-379, Rockland) shaking delicately at 4°C. Secondary antibodies (Alexa, 1:200, Jackson ImmunoResearch) were incubated for 2h at room temperature. Finally, slices were incubated with 40,6-diamidino-2-phenylindole solution (DAPI, 1:10,000, Invitrogen) for 10 min and mounted in Mowiol medium. Images were acquired with a confocal microscope (LSM700, Zeiss).

Rigter P.M., Wallaard I., Aghadavoud Jolfaei M., Kingma J., Post L., Elgersma M., Elgersma Y, & van Woerden G.M. (2022). Adult Camk2a gene reinstatement restores the learning and plasticity deficits of Camk2a knockout mice. iScience, 25(11), 105303.

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Immunohistochemistry of Kidney Sections

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Paraffin-embedded kidney sections were dewaxed, rehydrated, and antigen retrieval was carried out in microwave-boiled citrate buffer (10 mM citric acid, pH 6.0) for 1 h (fibronectin and Ki-67 staining). Sections were then permeabilized with 0.1% Triton X-100 in PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4) and blocked with blocking buffer [5% Normal goat serum, 0.1% Triton X-100 in PBS (PBS-TX)] for 1 h. Incubation with primary antibody diluted in blocking buffer is done at 4°C overnight. Anti-fibronectin (Abcam) at 1:250 dilution, anti-Ki-67 (Abcam) at 1:500 dilution, anti-Mfsd2a (in house) at 1:100, anti-RFP (Rockland) at 1:500 dilution was used. Sections are then washed three times the following day with PBS-TX and incubated with Alexa Fluor Plus secondary antibody (Thermo Fisher Scientific) diluted in blocking buffer (1:250) for 1 h. After washing three times with PBS-TX, Hoechst 33,342 (Thermo Fisher Scientific) was used to stain nuclei and the slides mounted with Fluorsave Reagent (Merck). Leica Fluorescence microscope (Leica Biosystems) was then used for image acquisition.

Loke R.Y., Chin C.F., Liang G., Wong B.H., Galam D.L., Tan B.C., Chua G.L., Minegishi S., Morisawa N., Sidorov I., Heijs B., Titze J., Wenk M.R., Torta F, & Silver D.L. (2023). Mfsd2a-mediated lysolipid transport is important for renal recovery after acute kidney injury. Journal of Lipid Research, 64(8), 100416.

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Immunoblot Analysis of Protein Expression

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Immunoblot analysis was performed as previously described (80 (link)). Briefly, cells were lysed in radioimmunoprecipitation assay buffer supplemented with a protease inhibitor cocktail (Roche). Whole-cell extracts were resolved by SDS–polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes, and probed with indicated primary antibodies. Bound antibodies were detected with horseradish peroxidase (HRP)–conjugated secondary antibodies and chemiluminescent HRP substrate. The following primary antibodies were used for Western blotting (all from CST, Beverly, MA, USA, unless otherwise indicated): anti-SOX9 (#82630; 1:1000), anti-vinculin (#13901; 1:1000), and anti-RFP (Rockland, 600-401-379; 1:500).

Bala P., Rennhack J.P., Aitymbayev D., Morris C., Moyer S.M., Duronio G.N., Doan P., Li Z., Liang X., Hornick J.L., Yurgelun M.B., Hahn W.C, & Sethi N.S. (2023). Aberrant cell state plasticity mediated by developmental reprogramming precedes colorectal cancer initiation. Science Advances, 9(13), eadf0927.

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Immunofluorescent Labeling of Pancreatic Glucagon

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Animals were anaesthetised with ketamine and transcardially perfused-fixed with 4% PFA before post-fixation for 1 h and embedding in paraffin and sectioning. Sections were labelled with anti-RFP (Rockland, 600-401-379, 1:50 dilution, secondary-Alexa 568 1:250) and anti-glucagon (1:500 dilution, secondary-Alexa 488 1:250) and anti-glucagon (1:500 dilution, secondary-Alexa 488 1:250) antibodies and sealed using Vectashield hardset mounting medium with DAPI (Vector Laboratories).

Soedling H., Hodson D.J., Adrianssens A.E., Gribble F.M., Reimann F., Trapp S, & Rutter G.A. (2015). Limited impact on glucose homeostasis of leptin receptor deletion from insulin- or proglucagon-expressing cells. Molecular Metabolism, 4(9), 619-630.

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Immunofluorescence Staining of Neural Progenitors

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Neural progenitor cells grown on poly-L-ornithine/laminin coated 4-well chamber plates were fixed in 4% paraformaldehyde for 15 min at room temperature, permeabilized with 0.3% Triton-X-100 in Triton Buffer Solution (TBS) for 10 min, blocked with 2% FBS in TBS-0.05% Tween-20 for 20 min and stained with primary antibodies in blocking buffer at 4°C overnight. Primary antibodies included: anti-Nestin (611658; BD Pharmingen); anti-SOX2 (561469; BD Pharmingen); anti-RFP (600-401-379; Rockland); anti-ki67-Alexa fluor-488 (51-9007231; BD Stemflow Human neural Lineage analysis Kit); TuJ1 (MMS-435P, Covance); GFAP (ab7779, Abcam). The next day, the cells were stained with secondary antibodies (Alexa fluor 568 donkey anti-rabbit; Alexa fluor 488 donkey anti-mouse; Life Technologies, Inc.) in blocking buffer for 45 min and nuclei were counterstained with DAPI. Images were acquired using an Olympus FV1000 confocal microscope. Live culture images were also acquired using an Inverted Axioscope and AxioCam MRm (Carl Zeiss, Inc.). Image assembly was performed using Adobe Photoshop CS5 (Adobe Systems, Inc.).

Fu X., Rong Z., Zhu S., Wang X., Xu Y, & Lake B.B. (2014). Genetic approach to track neural cell fate decisions using human embryonic stem cells. Protein & Cell, 5(1), 69-79.

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Dilution-based DREADD Delivery in Superior Colliculus

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To exclude retinal transduction, the DREADD viral vector was injected in the superior colliculus at different dilutions (1:2, 1:4 and 1:8 in PBS). Two weeks after stereotactic injection, mice were euthanized with an overdose of sodium pentobarbital (60 mg/kg, Dolethal, Vetoquinol, Aartselaar, Belgium) and transcardially perfused with 0.9% saline and 4% paraformaldehyde (PFA) in PBS. Retinas were dissected, flatmounted, and double stained with anti-Brn3a (1:750, Santa-Cruz Biotechnologies, Dallas, TX, USA) and anti-RFP (1:5000, Rockland, Philadelphia, PA, USA), as previously described [14 (link)].

Claes M., Geeraerts E., Plaisance S., Mentens S., Van den Haute C., De Groef L., Arckens L, & Moons L. (2022). Chronic Chemogenetic Activation of the Superior Colliculus in Glaucomatous Mice: Local and Retrograde Molecular Signature. Cells, 11(11), 1784.

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Antibody Characterization and Usage Protocol

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Anti-Rbfox1(A2BP1) was produced by ourselves as previously described28 (link). The following mouse monoclonal antibodies were used; anti-Tau-1 (MAB3420; Chemicon International, Temecula, CA), anti-Myc 9E10 and anti-MAP2 (M4403; Sigma-Aldrich, St Louis, MO). Polyclonal rabbit antibodies used were anti-GFP (#598; MBL, Nagoya, Japan), anti-RFP (#600-401-379; Rockland Immunochemicals, Gilbertsville, PA), anti-Rbfox2 (Fox2) (A300-864A-T, Bethyl Laboratories, Montgomery, TX) and anti-Sept1129 (link). anti-GFP (GFP-1020; Chicken) was purchased from AVES Labs (Tigard, OR).

Hamada N., Ito H., Nishijo T., Iwamoto I., Morishita R., Tabata H., Momiyama T, & Nagata K.I. (2016). Essential role of the nuclear isoform of RBFOX1, a candidate gene for autism spectrum disorders, in the brain development. Scientific Reports, 6, 30805.

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Zebrafish Ventricular Injury Proliferation Assay

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Zebrafish ventricles were extracted from adults that had not been treated with morphine and fixed in 4% PFA for 1 h at room temperature. Ventricles were embedded in OCT (Tissue-Tek) and sectioned (8 µm). For each biological replicate, three nonconsecutive midsagittal sections were used. Immunostaining was performed as previously described^{65 (link)}. The primary antibodies used were anti-RFP (Rockland, catalog no. 600-401-379, 1:200) and anti-PCNA (Santa Cruz Biotechnology, catalog no. sc-56, 1:500). The secondary antibodies were AlexaFluor 488 goat anti-mouse IgG (H+L) (Thermo Scientific, catalog no. A-11029, 1:500) and AlexaFluor 647 goat anti-rabbit IgG (H+L) (Thermo Scientific, catalog no. A-21244, 1:500). Confocal images were obtained using an LSM700 microscope (Zeiss). The percentage of proliferating cECs (PCNA⁺) was calculated as the ratio of the total numbers of cECs in the injury area and in 200 µm of the injury border zone using ZEN 3.2 Blue software. For fluorescence intensity analysis, the percentage fluorescence was calculated from whole-mount images as a ratio to the background fluorescence in the injury area using Fiji software.

Hu B., Lelek S., Spanjaard B., El-Sammak H., Simões M.G., Mintcheva J., Aliee H., Schäfer R., Meyer A.M., Theis F., Stainier D.Y., Panáková D, & Junker J.P. (2022). Origin and function of activated fibroblast states during zebrafish heart regeneration. Nature Genetics, 54(8), 1227-1237.

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Immunofluorescence Antibody Protocol

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Primary antibodies used in this project and their final concentrations were as follows: anti-GFP (Life technology, Carlsbad, CA, USA, 1:1000), anti-RFP (Rockland, Limerick, PA, USA, 1:1000), anti-β-actin (Sigma-Aldrich, St. Louis, MO, USA, 1:5000), anti-GM130 (BD, Franklin Lakes, NJ, USA, 1:500), anti-EEA1 (BD, Franklin Lakes, NJ, USA, 1:500), anti-Rab 7 (Santa Cruz, Santa Cruz, CA, USA, 1:500), anti-LAMP1 (DSHB, Iowa, IA, USA, 1:500), anti-PRR/ATP6AP2 (Sigma-Aldrich, St. Louis, MO, USA, 1:500), anti-APP (Cell Signaling Technology, Danvers, MA, USA, 1:500), anti-sAPPβ (a gift from Tae-Wan Kim, 1:500). All the corresponding conjugated secondary antibody (1:1000) were purchased from Invitrogen (Waltham, MA, USA). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (1:1000, Roche, Basel, Switzerland).

Zhao L., Zhao Y., Tang F.L., Xiong L., Su C., Mei L., Zhu X.J, & Xiong W.C. (2019). pHluorin-BACE1-mCherry Acts as a Reporter for the Intracellular Distribution of Active BACE1 In Vitro and In Vivo. Cells, 8(5), 474.

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Immunohistochemistry of Rodent Brain

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Animals were deeply anesthetized with sodium pentobarbital (nembutal; 80 mg/kg, i.p.) and perfused with PBS (0.1 M) and 4% paraformaldehyde in PBS (4% PFA/PBS, 4°C, 500 ml). The dissected brains were post-fixed at 4°C in 4% PFA/PBS and cryo-protected at 4°C in 30% sucrose/PBS. Coronal sections (50 μm) were prepared using a vibrating blade microtome (Leica, CM1950). All sections were post-fixed for 20 min at 4°C in 4% PFA/PBS. Sections were blocked and permeabilized for 1 hour at room temperature in a PBS solution containing 5% bovine serum albumin (BSA) and 0.3% Triton X-100. The primary antibody incubation was performed by incubating the sections overnight at 4°C in a PBS solution containing 5% BSA, polyclonal anti-RFP (1:500, Rockland, Cat. No. 600-401-379), and anti-PV (1:500, Swant, Cat. No. 235). The secondary antibody incubation was performed for 1 hr using Alexa Fluor 594 donkey anti-rabbit IgG and Alexa Fluor 488 donkey anti-mouse IgG (1:500, Thermo Fisher Cat. No. A32754 and Cat. No. A21202, respectively). Nuclei were stained with DAPI (Sigma-Aldrich, Cat. No. D9542) shown in the blue channel. Brain sections were mounted onto slides using Fluoromount-G mounting medium (SouthernBiotech, Cat. No. 0100–01).

Zhang Y., Roy D.S., Zhu Y., Chen Y., Aida T., Hou Y., Shen C., Lea N.E., Schroeder M.E., Skaggs K.M., Sullivan H.A., Fischer K.B., Callaway E.M., Wickersham I.R., Dai J., Li X.M., Lu Z, & Feng G. (2022). Targeting thalamic circuits rescues motor and mood deficits in PD mice. Nature, 607(7918), 321-329.

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