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Rabbit anti rfp

Manufactured by Rockland Immunochemicals
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Rabbit anti-RFP is a primary antibody that specifically binds to the red fluorescent protein (RFP) from various sources. It can be used for detection and localization of RFP-tagged proteins in various applications, such as Western blotting, immunohistochemistry, and immunocytochemistry.

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Lab products found in correlation

Chicken anti gfp, by Thermo Fisher Scientific (10 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (10 mentions) Lsm 710, by Zeiss (8 mentions) Chicken anti gfp, by Abcam (7 mentions) Rat anti ctip2, by Abcam (6 mentions) Chicken anti gfp, by Aves Labs (6 mentions) Alexa fluor 488 goat anti chicken, by Thermo Fisher Scientific (5 mentions) Mouse anti satb2, by Abcam (4 mentions) Alexa fluor 555 goat anti rabbit, by Thermo Fisher Scientific (4 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (4 mentions) Lsm800 confocal microscope, by Zeiss (4 mentions) Chick anti gfp, by Abcam (3 mentions) Mouse anti neun, by Merck Group (3 mentions) Cryostat, by Leica camera (3 mentions) Goat anti gfp, by Abcam (3 mentions) Goat anti rat alexa fluor 647, by Thermo Fisher Scientific (3 mentions) Donkey anti rabbit alexa 568, by Thermo Fisher Scientific (3 mentions) Axio observer z1, by Zeiss (3 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions)

91 protocols using rabbit anti rfp

1

Immunostaining and 3DISCO Protocols

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An overview of the antibodies used is presented in Table 1 and Table 2. For DAB staining, we used a primary rabbit anti-RFP antibody (1:1000, Rockland Immunochemicals, Limerick, PA, USA) and a secondary donkey anti-rabbit antibody (1:200, Jackson, Cambridge, UK). For immunofluorescence, we used chicken anti-calbindin (1:500, Synaptic Systems, Goettingen, Germany), mouse anti-calbindinD-28 (1:500, Sigma-Aldrich), chick anti-GFP (1:200, Abcam, Cambridge, UK), goat anti-FoxP2 (1:500, Santa Cruz, Santa Cruz, CA, USA), guinea pig anti-vGluT2 (1:500, MilliPore, Amsterdam, The Netherlands), rabbit anti-RFP (1:1000, Rockland) and mouse anti-NeuN (1:1000, MilliPore) as primary antibodies. Cy5 anti-chicken (1:200, Jackson), Cy3 anti-mouse, FITC anti-chick, (1:200, MilliPore), Alexa488 anti-goat (1:200, Jackson), Cy5 anti-guinea pig (1:200, Jackson), Cy3 anti-rabbit (1:400, Jackson) and Alexa488 anti-Mouse (1:200, Jackson) antibodies were used as secondary antibodies.
For 3DISCO, we used chicken anti-calbindin (1:500, Synaptic Systems), goat anti-FoxP2 (1:500, Santa Cruz) and rabbit anti-RFP (1:000, Rockland) as primary antibodies. Cy5 anti-chicken (1:200, Jackson), Alexa488 anti-goat (1:200, Jackson) and Cy3 anti-rabbit (1:400, Jackson) antibodies were used as secondary antibodies.
Dumas D.B., Gornati S.V., Adolfs Y., Shimogori T., Pasterkamp R.J, & Hoebeek F.E. (2022). Anatomical Development of the Cerebellothalamic Tract in Embryonic Mice. Cells, 11(23), 3800.
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2

Immunostaining of Engineered Cultured Neurons

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Cultured hippocampal neurons were created from WT P0 pups as described above. Neurons were infected at DIV0–DIV1 with AAV-U6-CARMIL3Ctermsg1-HITI-smFP-P2A-mCherry-SynI-Cre and also either infected with pAAV-pMecp2-SpCas9-spA (AAV-Cas9), PX551, or lipofectamine transfected at DIV2 with PX330 (Cas9). At DIV14–16, neurons were fixed in and prepped for primary antibody incubation as described above. Neurons were incubated in primary antibodies: Rat Anti-HA (Sigma, 3F10, 1:200), Chicken Anti-GFP (Abcam, ab13970, 1:1000), and Rabbit anti-RFP (Rockland, 600-041-379, 1:1000) for 16–18 h at 4 °C. Neurons were then washed three times in PBS for 5min each and incubated in secondary antibodies: Alexa Fluor 647 Goat anti-Rat (ThermoFisher, A-21247, 1:1000), Alexa Fluor 488 Goat anti-Chicken (ThermoFisher, A-11006, 1:1000), and Alexa Fluor 555 Goat anti-Rabbit (ThermoFisher, A32732, 1:1000) for 1–2 h. Coverslips were then mounted with mounting media (Fluorsave Reagent, EMD Millipore, 345789) and imaged on a Zeiss 880 Airyscan inverted confocal microscope.
Spence E.F., Dube S., Uezu A., Locke M., Soderblom E.J, & Soderling S.H. (2019). In vivo proximity proteomics of nascent synapses reveals a novel regulator of cytoskeleton-mediated synaptic maturation. Nature Communications, 10, 386.
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3

Immunofluorescent Staining of Transplanted Cells

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Subsequent to the electrophysiology recordings, the slices were fixed overnight in 4% paraformaldehyde (Electron Microscopy Sciences) in 0.1 M phosphate buffer (pH 7.4). The transplanted cells were identified in the slices by immunofluorescent staining for human nuclear antigen (mouse anti-HuNu, 1:1000, Chemicon) or the expression of RFP (rabbit anti-RFP 1:1000, Rockland,). Biocytin staining was performed by incubating the slices in Streptavidin 647 (1:1000, Life Technologies) [14 (link)].
Shrestha S., Anderson N.C., Grabel L.B., Naegele J.R, & Aaron G.B. (2020). Development of electrophysiological and morphological properties of human embryonic stem cell-derived GABAergic interneurons at different times after transplantation into the mouse hippocampus. PLoS ONE, 15(8), e0237426.
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4

Immunofluorescence Staining of Drosophila Tissues

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All antibodies that are used in this study are listed in the Key resources table. The following primary antibodies were used: rat-anti-E-Cad (1:200, DHSB DCAD2 concentrate), mouse-anti-β-catenin (1:400, DHSB N2 7A1 Armadillo concentrate), mouse-anti-LacZ (1:100, DHSB 40-1a concentrate), rat-anti-HA (1:100, Anti-HA High Affinity rat IgG1, Roche ROAHAHA). Sqh::eGFP was detected with rabbit-anti-GFP (1:500, Life Technologies A11122 or 1:1000 Abcam ab6556). Cirl::RFP was detected with rabbit-anti-RFP (1:1000, Rockland 600-401-379). The following secondary antibodies were used: donkey-anti-rabbit Alexa Fluor 488 IgG (Life Technologies A 21206), donkey-anti-rabbit Alexa Fluor 568 IgG (Life Technologies A10042), donkey anti-mouse Alexa Fluor 568 IgG (Life Technologies A10037), donkey-anti-mouse Alexa Fluor 647 IgG (Jackson ImmunoResearch 715 605 151) and donkey-anti-rat Alexa Fluor 647 IgG (Jackson ImmunoResearch 712 605 153). All secondary antibodies were used at 1:500.
Lavalou J., Mao Q., Harmansa S., Kerridge S., Lellouch A.C., Philippe J.M., Audebert S., Camoin L, & Lecuit T. (2021). Formation of polarized contractile interfaces by self-organized Toll-8/Cirl GPCR asymmetry. Developmental Cell, 56(11), 1574-1588.e7.
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5

Western Blot Analysis of T. gondii Proteins

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T. gondii parasites were harvested (1 × 106), lysed in radioimmunoprecipitation assay (RIPA) lysis buffer, boiled in 1× NuPAGE LDS sample buffer, loaded onto precast 10% Any-KD Mini-Protean TGX gels (Bio-Rad), and run at 150 V (13 (link)). Proteins were transferred to nitrocellulose membranes and probed with antibodies mouse anti-PUBL (generated in this study) at 1:200, rabbit anti-CDC48AP (14 (link)) at 1:500, mouse anti-tubulin (12G10; a gift from Jacek Gaertig, University of Georgia) at 1:2,000, rat anti-HA (clone 3F10; Roche Applied Science) at 1:400, mouse anti-GFP (Torry Pines Biolab) at 1:500, and rabbit anti-RFP (Rockland Immunochemicals) at 1:1,000 followed by incubation of the membranes with horseradish peroxidase-conjugated anti-mouse, anti-rat, or anti-rabbit antibodies (Bio-Rad) (1:10,000 dilution). Bands were detected by incubation of the membrane with Pierce ECL Western blotting substrate and exposure of the membrane to film.
Fellows J.D., Cipriano M.J., Agrawal S, & Striepen B. (2017). A Plastid Protein That Evolved from Ubiquitin and Is Required for Apicoplast Protein Import in Toxoplasma gondii. mBio, 8(3), e00950-17.
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6

Immunofluorescence Staining of Ileum Tissue

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For staining, terminal ileum tissue was fixed in 10% formalin and embedded in paraffin blocks. Paraffin-embedded tissue sections (5 μm) were deparaffinized using Xylene and rehydrated using a descending ethanol gradient. Antigen retrieval was conducted by heating slides in a microwave. Tissue sections were blocked in 5% bovine serum albumin in 1× PBS for 1 h at 37 °C. Tissue sections were incubated at 4 °C overnight in primary antibodies against LYZ1-FITC (Dako, 1:200), MMP7 (Cell Signaling Technology, 1:200). Slides were washed three times for 5 min, and sections were incubated with anti-rabbit IgG Fab2 Alexa Fluor® 647 (Cell Signaling Technology, 1:300). Slides were mounted using a DAPI hard stain (Vector® Laboratories, H-1500) to visualize cell nuclei. Images were acquired using a Zeiss 510 Meta NLO confocal microscope (Stony Brook University).
For lineage-tracing experiments, Rabbit anti-RFP (Rockland, 1:300) was used as a primary antibody, Mouse anti-rabbit (Jackson Laboratories, 1:300) was used as a secondary antibody and Donkey anti-mouse-Cy5 (Jackson Laboratories, 1:300) was used as a tertiary antibody. The primary antibody was applied overnight at 4 °C, and the secondary and tertiary antibodies were applied for 30 min at 37 °C. Slides were washed three times for 5 min after each antibody was applied.
Gaudino S.J., Beaupre M., Lin X., Joshi P., Rathi S., McLaughlin P.A., Kempen C., Mehta N., Eskiocak O., Yueh B., Blumberg R.S., van der Velden A.W., Shroyer K.R., Bialkowska A.B., Beyaz S, & Kumar P. (2020). IL-22 receptor signaling in Paneth cells is critical for their maturation, microbiota colonization, Th17-related immune responses, and anti-Salmonella immunity. Mucosal Immunology, 14(2), 389-401.
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7

Whole-Brain Immunolabeling and Clearing Protocol

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Mouse brains were cleared and immunolabeled following the procedures decribed by Renier et al. (2016) . Samples were processed with the May 2016 version of the iDISCO+ protocol available at http://idisco.info. Briefly, n = 4 adult male PnocIRES-Cre; Ai9 reporter mice with deeply anesthetized and transcardially perfused with 30mL of PBS followed by 30mL of 4% PFA in PBS. Samples were post-fixed overnight at 4°C in 4% PFA in PBS and then stored in PBS at 4°C until use. Individual brains were hemisected and the hemisphere taken from each animal (left or right) was counterbalanced to include an equal number of hemispheres from each side. Brains then underwent sample pretreatment, permeabilization, and blocking as described in the protocol. Hemispheres were then incubated for 7 days at 37°C with shaking in 1:1000 Rabbit anti-RFP (Rockland Immunochemicals #600–401-379) recognizing tdTomato in the reporter mice. Samples then underwent 1 day of washing as described in the protocol and were incubated for 7 days at 37°C with shaking in 1:500 Donkey anti-Rabbit Alexa Fluor 647 (Thermo Fisher #A-31573). Samples then underwent one final wash followed by dehydration and tissue clearing as described in the protocol. Brains were stored in the dark at room temperature in Dibenzyl ether until they were imaged.
Hardaway J.A., Halladay L.R., Mazzone C.M., Pati D., Bloodgood D.W., Kim M., Jensen J., DiBerto J.F., Boyt K.M., Shiddapur A., Erfani A., Hon O.J., Neira S., Stanhope C.M., Sugam J.A., Saddoris M.P., Tipton G., McElligott Z., Jhou T.C., Stuber G.D., Bruchas M.R., Bulik C.M., Holmes A, & Kash T.L. (2019). Central Amygdala Prepronociceptin-Expressing Neurons Mediate Palatable Food Consumption and Reward. Neuron, 102(5), 1037-1052.e7.
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8

Drosophila Tissue Immunostaining Protocol

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The primary antibodies used were as follows: anti-DIP-γ (guinea pig, 1:200) and anti-Dpr11 (rabbit, 1:150) were gifts from C. Desplan. Anti-Pros MR1A (mouse 1:4), anti-Elav 7E8A10 (rat, 1:10), anti-Dac 2–3 (mouse 1:50), anti-chaoptin 24B10 (mouse 1:20) were obtained from Developmental Studies Hybridoma Bank (University of Iowa, IA). Commercial antibodies were used as follows: Rabbit anti-RFP (Rockland, 1:500), rabbit anti-GFP (Thermo- Fisher Scientific, 1:500), chicken anti-GFP (Aves labs, 1:500), mouse anti-myc 9E10 (Abcam, 1:500) and chicken anti-beta-galactosidase (Abcam, 1:1000). Secondary antibodies were obtained from Thermo-Fisher Scientific and used at 1:500.
Menon K.P., Kulkarni V., Takemura S.Y., Anaya M, & Zinn K. (2019). Interactions between Dpr11 and DIP-γ control selection of amacrine neurons in Drosophila color vision circuits. eLife, 8, e48935.
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9

Immunostaining of Adult Fly Brains

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Dissection and immunostaining of adult fly brains was performed as previously described [12 (link)]. The primary antibodies employed were rabbit anti-GFP 1:500 (Invitrogen), rabbit anti-RFP 1:500 (Rockland), chicken anti-GFP 1:500 (Upstate), rabbit anti-PDF 1:1500 (custom-made by NeoMPS, France) and homemade rat anti-Drosophila-PDF 1:500 [12 (link)]. Secondary antibodies used were Cy2-and Cy3-conjugated anti-rabbit, Cy2-conjugated anti-chicken and Cy5- and Cy3-conjugated anti-rat (Jackson ImmunoResearch). Images were taken on either a Zeiss Pascal LSM, a Zeiss LSM 510 Meta confocal or a Zeiss LSM 710 two-photon microscope. After acquisition, images were processed employing Fiji, an ImageJ-based image processing environment [44 (link)].
Gorostiza E.A., Depetris-Chauvin A., Frenkel L., Pírez N, & Ceriani M.F. (2014). Circadian pacemaker neurons change synaptic contacts across the day. Current biology : CB, 24(18), 2161-2167.
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10

Immunohistochemical Analysis of Brain Samples

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Immunohistochemistry follows protocols previously reported15 (link),16 ,18 (link). Mice were overdosed with 3% isoflurane and perfused transcardially with cold (4 °C) phosphate-buffered saline (PBS) followed by 4% paraformaldehyde (PFA) in PBS. Brains were extracted and stored overnight in PFA at 4 °C. Fifty μm coronal sections were collected in serial order using a vibratome and collected in cold PBS. Sections were blocked for 1 hour at room temperature in PBST and 5% normal goat serum (NGS) or Bovine albumin serum (BSA) on a shaker. Sections were transferred to wells containing primary antibodies (1:1000 guinea anti-c-Fos [SySy]; 1:1000 rabbit anti-RFP [Rockland]; 1:5000 chicken anti-GFP [Invitrogen]) and allowed to incubate on a shaker overnight at room temperature or 3 days at 4 degrees C. Sections were then washed in PBST for 10-min (x3), followed by 2-hour incubation with secondary antibody (1:200 Alexa 555 anti-rabbit [Invitrogen]; 1:200 Alexa anti-guinea 647 [Invitrogen]; 1:200 Alexa 488 anti-chicken [Invitrogen]). Following three additional 10-min washes in PBST, sections were mounted onto micro slides (VWR International, LLC). Vectashield Hart Set Mounting Medium with DAPI (Vector Laboratories, Inc) was applied, slides were coverslipped, and allowed to dry overnight.
Shpokayte M., McKissick O., Guan X., Yuan B., Rahsepar B., Fernandez F.R., Ruesch E., Grella S.L., White J.A., Liu X.S, & Ramirez S. (2022). Hippocampal cells segregate positive and negative engrams. Communications Biology, 5, 1009.
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