Rabbit anti rfp

Manufactured by Abcam

Sourced in United States, United Kingdom

Rabbit anti-RFP is a primary antibody that specifically recognizes and binds to the red fluorescent protein (RFP) tag. It is commonly used in various research applications, such as immunohistochemistry, immunocytochemistry, and Western blotting, to detect and visualize RFP-tagged proteins in biological samples.

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Lab products found in correlation

Chicken anti gfp, by Abcam (7 mentions) Rabbit anti ha, by Cell Signaling Technology (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions) Rabbit anti apkc, by Santa Cruz Biotechnology (2 mentions) Guinea pig anti dcx, by Merck Group (2 mentions) Goat anti c fos, by Santa Cruz Biotechnology (2 mentions) Mouse anti brdu, by BD (2 mentions) Mouse anti neun, by Merck Group (2 mentions) Triton x 100, by Merck Group (2 mentions) Fluoview 1000, by Olympus (2 mentions) Mouse anti mira, by Abcam (2 mentions) Rat anti ph3, by Abcam (2 mentions) Chick anti gfp, by Abcam (2 mentions) Mouse anti gfp, by Abcam (2 mentions) Mouse monoclonal nc82, by Developmental Studies Hybridoma Bank (2 mentions) Goat anti mouse cy5, by Jackson ImmunoResearch (2 mentions) Goat anti chicken alexa488, by Abcam (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Confocal microscope, by Zeiss (1 mentions) Sheep anti gfp, by Bio-Rad (1 mentions)

Close spelling variants of this product

Rabbit anti-RFP antibody (4 citations) Anti-RFP (19 citations) Rabbit anti- (5 citations)

22 protocols using rabbit anti rfp

Immunostaining of Drosophila Neuronal Tissues

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Antibodies used in this work include Guinea pig anti Oc (1:750, gift from Tiffany Cook) (Xie et al., 2007 (link)); Rabbit anti Sox102F and Rabbit anti Slp1 (1:500 for both, Gifts from Claude Desplan); Rabbit anti-D (1:200) (from John R. Nambu), mouse anti-Ey (1:10, DSHB), sheep anti-GFP (1:500, AbD Serotec), Goat anti anti-beta-gal (Abcam 1:1000), rabbit anti-RFP (Abcam 1:1000), Rabbit anti HA (Cell Signaling Technology, 1:1000). Secondary antibodies are from Jackson or Invitrogen. Immunostaining was done as described (Li et al., 2013 (link)) with a few modifications: 3^rd instar Larval brains or adult brains were dissected in 1XPBS, and fixed in 4% Formaldehyde for 30 minutes on ice (larval) or 45min at RT (adult). Brains were incubated in primary antibody solution overnight at 4°C, washed three times and incubated in secondary antibody solution overnight at 4°C, washed three times and mounted in Slowfade. Images are acquired using a Zeiss Confocal Microscope. Figures are assembled using Photoshop and Illustrator.

Naidu V.G., Zhang Y., Lowe S., Ray A., Zhu H, & Li X. (2020). Temporal progression of Drosophila medulla neuroblasts generates the transcription factor combination to control T1 neuron morphogenesis. Developmental biology, 464(1), 35-44.

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Immunofluorescence Staining for Angiogenesis

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Immunofluorescence staining was performed according to the protocol previously described [28 (link)]. The primary antibodies used were as follows: rabbit anti-RFP (1:200; Abcam Inc., USA), mouse anti-PECAM-1 (1:50; Santa Cruz Biotechnology, Germany), rabbit anti-Ki67 (1:200; Cell signaling technology, USA). Each section was washed with PBS and incubated with proper secondary antibodies at room temperature for 2 h the following day. 1%BSA was used as a control to confirm the specificity of the antibody and DAPI (1:30) was used to detect the nucleus. Images were acquired using a Leica fluorescence microscope (Leica, Germany). Ki67⁺ /CD31⁺ microvessels in the perifocal region was counted and quantified by an investigator who was blinded to the experimental groups.

Liu J., Li Q., Zhang K.S., Hu B., Niu X., Zhou S.M., Li S.G., Luo Y.P., Wang Y, & Deng Z.F. (2016). Downregulation of the Long Non-Coding RNA Meg3 Promotes Angiogenesis After Ischemic Brain Injury by Activating Notch Signaling. Molecular Neurobiology, 54(10), 8179-8190.

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Tissue Fixation and Immunolabeling Protocol

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Mice were transcardially perfused using 4% paraformaldehyde (PFA) prepared in phosphate buffered saline (PBS) and tissues harvested were postfixed in 4% PFA overnight. Brains were sagittally sectioned to 50um thick sections using vibratome (Leica VT 1000S) and stored in a solution of 1xPBS with 0.05% sodium azide. Similarly lungs, testis, and oviducts were sectioned coronally. For immunohistochemical analysis tissue sections were immersed in blocking buffer (10% goat serum and 1% Triton-X-100 in 1xPBS) for an hour at room temperature (RT). Following blocking, sections were incubated with appropriate primary antibodies [chicken anti-GFP (Abcam) at 1:2000; rabbit anti-RFP (Abcam) at 1:1000; mouse anti-Tubulin (Sigma) at 1:1000; rabbit S100 (DAKO) at 1:1000] in antibody diluent (1% goat serum and 0.03% Triton-X-100 in 1xPBS) for overnight at 4°C. Sections were then washed three times, 5 minutes each, using 1xPBS and incubated with appropriate secondary antibodies in antibody diluent with DAPI [(Sigma) at 1µg/ mL] for an hour at RT. Following PBS washes, sections were mounted to plus-sided slides and coverslip placed on sections with mounting media to avoid air bubbles. Confocal images of stained sections were captured using either Nikon Eclipse C1 or Olympus fluoview FV1000 confocal microscopes.

Muthusamy N., Vijayakumar A., Cheng G J.r, & Ghashghaei H.T. (2014). A knock-in Foxj1CreERT2::GFP mouse for recombination in epithelial cells with motile cilia. Genesis (New York, N.Y. : 2000), 52(4), 350-358.

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Immunostaining of Transgenic Mouse Brains

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Animals were sacrificed 3 months after birth by terminal transcardial perfusion of 4% paraformaldehyde (PFA, Electron Microscopy Sciences) followed by overnight post-fixation in 4% PFA. Brains were washed 3X with PBS for 5 min and sectioned at 100-μm thickness using vibratome (Leica). Sections were permeabilized with 0.2% Triton X-100 3X for 5 min, blocked in PBS-based blocking buffer with 5% BSA and 0.2% Triton, and stained with chicken anti-GFP (Aves, 1:1000) and rabbit anti-RFP (Abcam, 1:1000) for enhancement of PSCV2-mVenus and pCAG-mito-DsRed, respectively overnight at 4 °C. Sections were washed with 0.2% Triton buffer 3X for 5 min and incubated with Alexa488- and Alexa 555- labeled secondary antibodies for PSCV2-mVenus and pCAG-mito-DsRed, respectively overnight at 4 °C. Sections were washed with 0.2% Triton buffer 3X for 10 min each and mounted using VectaShield® Mounting Medium (Vector Laboratory).

Lee A., Kondapalli C., Virga D.M., Lewis TL J.r., Koo S.Y., Ashok A., Mairet-Coello G., Herzig S., Foretz M., Viollet B., Shaw R., Sproul A, & Polleux F. (2022). Aβ42 oligomers trigger synaptic loss through CAMKK2-AMPK-dependent effectors coordinating mitochondrial fission and mitophagy. Nature Communications, 13, 4444.

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Immunohistochemical Analysis of Pupal Wings

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Pupal wings were fixed in 3.7% formaldehyde (Sigma-Aldrich) at 4°C overnight. Wing imaginal discs were fixed in 3.7% formaldehyde at room temperature (RT) for 20 minutes. All immunostaining and in situ hybridizations were performed as described previously [7 (link),9 (link)]. The primary antibodies used are as follows: mouse anti-DLG1, rat anti-DE-Cadherin and mouse anti-GFP (for immunohistochemistry; all at 1:50) were obtained from Developmental Studies Hybridoma Bank, rabbit anti-phospho-SMAD1/5 (1: 200 for IF, 1:2000 for Western blotting) from Cell Signaling Technology (CST), rabbit anti-Rab5 (1:600) and rabbit anti-RFP (1:5000 for Western blotting) from Abcam, mouse anti-RFP (1:5000 for Western blotting) from Chromotek, mouse anti-GFP (1: 5000 for Western blotting) from Millipore, mouse anti-β-tubulin (1:5000) from Sigma-Aldrich, rabbit anti-MYC (1:500), goat anti-Scrib (1:100), rabbit anti-aPKC (1:100) and mouse anti-LGL (1:200) from Santa Cruz Biotechnology, and rabbit anti-Scrib (1:2000) from C. Doe. Secondary antibodies were as follows: goat anti-mouse IgG Alexa 488, goat anti-mouse IgG Alexa 568, goat anti-mouse IgG Alexa 647, goat anti-rabbit IgG Alexa 568, goat anti-rabbit IgG Alexa 647, goat anti-rat IgG Alexa 488 and goat anti-mouse IgG Cy5, all from Molecular Probes (1:200). GFP-booster (1:200, ChromoTek) was used to enhance the YFP signal in Fig 3F and S2C Fig.

Gui J., Huang Y, & Shimmi O. (2016). Scribbled Optimizes BMP Signaling through Its Receptor Internalization to the Rab5 Endosome and Promote Robust Epithelial Morphogenesis. PLoS Genetics, 12(11), e1006424.

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Multimodal Immunohistochemistry for Neural Phenotypes

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Mice were intracardially perfused with 4% paraformaldehyde (PFA; in 0.1 M PBS; pH 7.4) 90 minutes after the last testing session, and brains were harvested and post-fixed overnight in 4% PFA at 4°C. Brains were sectioned on a vibratome at 50 μm in the sagittal plane, blocked in 0.1 M PBS containing 10% serum and 1% Triton-X-100 (Sigma) for one hour at room temperature. For staining of BrdU, sections were washed in 2 N HCl for 10 minutes followed by three 5 minute washes in 0.1 M Sodium Borate (pH 8.5), followed by blocking. Sections were incubated with primary antibodies in 0.1 M PBS containing 1% serum and 0.3% Triton-X-100 overnight at 4°C, followed by washing in PBS and incubation with appropriate secondary antibodies for 1–2 hours at room temperature for visualization the next day. Antibodies and stains used: goat anti-cFos (Santa Cruz; 1:1000), chicken anti-GFP (Abcam; 1:2000), rabbit anti-RFP (Abcam; 1:1000), rabbit anti-HA (Cell Signaling; 1:500), mouse anti-NeuN (Millipore; 1:1000), mouse anti-BrdU (Becton Dickinson, 13:1000), guinea pig anti-Dcx (Millipore, 1:1000), and DAPI (nuclear stain; Sigma; 1:2000). Labeled sections were imaged on Fluoview 1000 (Olympus) or C1 (Nikon) confocal microscopes and double labeling was confirmed by z-stack analyses in the IMARIS software.

Muthusamy N., Zhang X., Johnson C.A., Yadav P.N, & Ghashghaei H.T. (2016). Developmentally defined forebrain circuits regulate appetitive and aversive olfactory learning. Nature neuroscience, 20(1), 20-23.

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Multimodal Immunohistochemistry Assay

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Immunohistochemistry was performed on free-floating sections. When antigen retrieval was necessary, tissues were treated for 45 minutes in citrate buffer (pH 6) at 80°C. To block unspecific staining when mouse-raised primary antibodies were used, we initially incubated sections with goat anti-mouse and goat anti-rabbit IgG (Jackson ImmunoResearch) 1:50 in 0.1 M PB 0.4% Triton-X (PBTx). After a permeabilization and blocking incubation for 2 hours at RT in PBTx supplemented with 10% heat-inactivated GS, the sections were incubated with the primary antibodies for 12 hours at 4°C in PBTx, 2% GS. Incubation with secondary antibodies was performed for 2 hours at 4°C in PBTx, 2% GS. Nuclear counterstaining was achieved with DAPI (0.5 μg/mL). The following primary antibodies and dilutions were used: mouse anti–Ki67 (1:100, Dako), rabbit anti-RFP (1:100, Abcam), rabbit anti-Sox2 (Cell Signaling, 1:100). For blood vessel staining, sections were incubated with DyLight-488 Tomato Lectin (Vector Lab) for 1hr RT at 1:1000. Coverslips were mounted using Pro-long Gold (Vector Lab).

Truong D., Fiorelli R., Barrientos E.S., Melendez E.L., Sanai N., Mehta S, & Nikkhah M. (2018). A Three-Dimensional (3D) Organotypic Microfluidic Model for Glioma Stem Cells – Vascular Interactions. Biomaterials, 198, 63-77.

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Immunostaining and FISH of Invertebrate Brains

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Immunostaining of embryonic, larval and adult brains were performed according to the described protocol (Büscher et al., 2019 (link); Hunnekuhl et al., 2019 (link)). FISH was performed using a horseradish peroxidase (POD) mediated tyramide signal amplification (TSA). Primary antibodies: chicken anti-GFP (1:1000, Abcam; RRID:AB_300798), mouse anti-acetylated Tubulin (1:50, Sigma; RRID:AB_609894), mouse anti-Synapsin (1:40, DHSB Hybridoma Bank; RRID:AB_2313867), rabbit anti-RFP (1:1000, Abcam; RRID: AB_945213), polyclonal rabbit anti-Tenascin (Teneurin)-a (Fascetti and Baumgartner, 2002 (link)). Secondary antibodies coupled with Alexa Fluor 488 or Alexa Fluor 555 (Thermo Fisher Scientific, RRID:AB_2633275 RRID:AB_2633276 RRID:AB_142924 RRID:AB_2535858) were used at 1:1000.

He B., Buescher M., Farnworth M.S., Strobl F., Stelzer E.H., Koniszewski N.D., Muehlen D, & Bucher G. (2019). An ancestral apical brain region contributes to the central complex under the control of foxQ2 in the beetle Tribolium. eLife, 8, e49065.

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Metastasis Quantification via Immunofluorescence

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Staining was conducted as previously published [11 (link), 22 (link)]. RFP staining: rabbit anti-RFP (Abcam, Cambridge, UK) 1:30 dilution, Alexa Fluor® 488 goat anti-rabbit secondary (Invitrogen); CDH1 staining: rabbit E-cadherin antibody (24E10; Cell Signaling Technology, Boston, MA, USA) 1:400 dilution. Internal negative controls were exposed to rabbit IgG or 10% goat serum rather than primary antibody. Images were captured on a Nikon TE2000 inverted microscope with IPLab software (BD Biosciences, Rockville, MD, USA), original magnification at 200× (RPF) or 400× (CDH1).
Metastases were calculated as a ratio of the number of RFP-positive cells versus the total amount of cells (determined by DAPI nuclear stain) per field of view. Points represent the ratio of RPF-positive cells versus DAPI for each section examined with mean for each group represented as horizontal black bar. SEM for each group indicated in red.

Rhodes L.V., Tate C.R., Segar H.C., Burks H.E., Phamduy T.B., Hoang V., Elliott S., Gilliam D., Pounder F.N., Anbalagan M., Chrisey D.B., Rowan B.G., Burow M.E, & Collins-Burow B.M. (2014). Suppression of triple-negative breast cancer metastasis by pan-DAC inhibitor panobinostat via inhibition of ZEB family of EMT master regulators. Breast cancer research and treatment, 145(3), 593-604.

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Immunostaining of Larval Drosophila Tissues

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Larval tissues were fixed for 20 min in 4% formaldehyde/phosphate-buffered saline (PBS) and immunostained as described (Bello et al. 2003 (link)). The primary antibodies used were rabbit anti-aPKC (1:500; Santa Cruz Biotechnology), mouse anti-Mira (1:50; gift of A. Gould), rat anti-pH3 (1:500; Abcam), chick anti-GFP (1:2000; Abcam), rabbit anti-Dpn (1:100; gift of Y.N. Jan), guinea pig anti-Dpn (1:1000; gift of James Skeath), rat anti-Pros (1:50; gift of F. Matsuzaki), rabbit anti-Ase (1:50; gift of F. Matsuzaki), rat anti-CycE (1:500; gift of Helena Richardson), mouse anti-CycA (1:50; Developmental Studies Hybridoma Bank), mouse or rat anti-Elav (1:100; Developmental Studies Hybridoma Bank), rabbit anti-Insc (1:1000; gift of W. Chia), rabbit anti-Myc (1:100; Santa Cruz Biotechnology), guinea pig anti-Nerfin-1 (1:5000; gift of A. Kuzin), rabbit anti-RFP (1:100; Abcam), and mouse anti-Fib (1:200; Abcam). Secondary goat antibodies conjugated to Alexa488, Alexa568, Alexa650, and Alexa505 (Molecular Probes) were used at 1:200. Images were collected on a Leica SP5 confocal microscope, and all images shown are single sections unless otherwise stated.

Froldi F., Szuperak M., Weng C.F., Shi W., Papenfuss A.T, & Cheng L.Y. (2015). The transcription factor Nerfin-1 prevents reversion of neurons into neural stem cells. Genes & Development, 29(2), 129-143.

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