Irdye 800cw goat anti rat igg

Manufactured by LI COR

The IRDye 800CW goat anti-rat IgG is a near-infrared fluorescent labeled secondary antibody. It is designed to detect and quantify rat immunoglobulin G (IgG) in various applications.

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Lab products found in correlation

Irdye 800cw donkey anti goat igg, by LI COR (4 mentions) High affinity anti ha rat and anti ha affinity matrix, by Roche (2 mentions) Irdye 800cw donkey anti rabbit igg, by LI COR (2 mentions) Irdye 800cw goat anti mouse igg, by LI COR (2 mentions) Irdye 800cw goat anti rabbit igg, by LI COR (2 mentions) Starbright blue 700 goat anti rabbit igg, by Bio-Rad (2 mentions) Fam83g, by Abcam (2 mentions) Starbright blue 700 goat anti mouse igg, by Bio-Rad (2 mentions) Lamin a c, by Cell Signaling Technology (2 mentions) α tubulin, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 goat anti rabbit igg, by Abcam (1 mentions) Ab52865, by Abcam (1 mentions) Ab90103, by Abcam (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Ab6046, by Abcam (1 mentions) Image studio, by LI COR (1 mentions) Irdye 680rd goat anti rabbit, by LI COR (1 mentions) Alpha tubulin, by Cell Signaling Technology (1 mentions) Ab8245, by Abcam (1 mentions) H3663, by Merck Group (1 mentions)

Close spelling variants of this product

IRDye 800CW goat anti-Rat (10 citations) Anti-rat IgG IRDye 800CW (2 citations) Anti-goat IRDye 800CW (5 citations) IgG IRDye 800CW (3 citations) IRDye 800CW (808 citations)

10 protocols using irdye 800cw goat anti rat igg

Phospho-specific GPR35 Antibody Characterization

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The rabbit phospho-site–specific GPR35 antiserum pSer³⁰⁰/pSer³⁰³-hGPR35a (Cat number (7TM0102C), raised against the sequence KAHKpSQDpSLCVTL, and the pSer²⁹⁸/pSer³⁰¹-mGPR35 antiserum (7TM0102B), raised against the sequence TPHKpSQDpSQILSLT, were developed in collaboration with 7TM Antibodies GmbH. IRDye 800CW donkey anti-rabbit IgG, IRDye 800CW donkey anti-goat IgG, and IRDye 800CW goat anti-rat IgG were from LI-COR Biosciences. Alexa Fluor 488-goat anti-rabbit IgG and Alexa Fluor 594-donkey anti-rat IgG were from Abcam. Mouse monoclonal anti-FLAG M2 was from Merck. Horseradish peroxidase anti-mouse (sheep) was from GE Healthcare. High affinity anti-HA (rat) and anti-HA affinity matrix were from Roche Diagnostics.

Divorty N., Jenkins L., Ganguly A., Butcher A.J., Hudson B.D., Schulz S., Tobin A.B., Nicklin S.A, & Milligan G. (2022). Agonist-induced phosphorylation of orthologues of the orphan receptor GPR35 functions as an activation sensor. The Journal of Biological Chemistry, 298(3), 101655.

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Western Blot Analysis of MSI1 Protein

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For mouse samples, 1–10 µg of protein per well was run on a 10% SDS gel. After transfer and blocking with 5% milk in TBST, immunoblotting was performed using the following primary antibodies: MSI1 (rat, 1:200, eBioscience #14H1), alpha-tubulin (rabbit, 1:1000, Cell Signaling #11H10). IRDye® 800CW goat anti-rat IgG (1:5000), IRDye® 680RD goat anti-rabbit (1:10,000) and IRDye® 800CW goat anti-mouse IgG (1:5000) were employed as the secondary antibodies (LICOR). All antibodies were diluted in 5% milk in TBST.
For human samples, denatured total cell protein (10 μg) was separated using 10% Bis-Tris gel electrophoresis and transferred to nitrocellulose membranes. Western blots were probed with the following antibodies: β-tubulin (rabbit; 1:50,000; Abcam #ab6046), GAPDH (mouse; 1:2000; Abcam #ab8245), MSI1 (rabbit; 1:2000; Abcam #ab52865) HIPK1 (1:500; Abcam #ab90103) (Table 1), Horseradish peroxidase conjugated with goat anti-rabbit IgG (1:20,000) or Licor anti-mouse (800 channel)/rabbit (700 channel) IgG were employed as the secondary antibodies (Bio-Rad). The bands were visualized with ImageStudio (Licor) or Chemidoc^TM MP Imaging Systems (Bio-Rad) using the ImageLab version 15.2.1 software.

Kameda-Smith M.M., Zhu H., Luo E.C., Suk Y., Xella A., Yee B., Chokshi C., Xing S., Tan F., Fox R.G., Adile A.A., Bakhshinyan D., Brown K., Gwynne W.D., Subapanditha M., Miletic P., Picard D., Burns I., Moffat J., Paruch K., Fleming A., Hope K., Provias J.P., Remke M., Lu Y., Reya T., Venugopal C., Reimand J., Wechsler-Reya R.J., Yeo G.W, & Singh S.K. (2022). Characterization of an RNA binding protein interactome reveals a context-specific post-transcriptional landscape of MYC-amplified medulloblastoma. Nature Communications, 13, 7506.

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Western Blot Analysis of Drosophila Proteins

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Whole fly lysates were prepared by homogenizing the adult flies in RIPA buffer (50 mM Tris-HCl [pH 7.4], 150 mM NaCl, 1% [v/v] NP-40, 0.1% [w/v] SDS, 0.5% [w/v] sodium deoxycholate,1 mM EDTA, 5 mM DTT, and 0.5 mM PMSF). The homogenates were clarified by centrifugation at 21,000g at 4°C for 10 min, and the supernatant was used for Western blot. Ovary lysates and the immunoprecipitation samples were prepared as above. Mouse anti-HA (Sigma, H3663), rabbit anti-Tubulin (Sigma, T3526), rabbit anti-α-Tubulin (Abcam, ab52866), rat anti-Vasa (Developmental Studies Hybridoma Bank), mouse anti-Dicer-2 (Miyoshi et al. 2009 (link)), mouse anti-R2D2 (Nishida et al. 2013 (link)), mouse anti-Ago2 (Miyoshi et al. 2005 (link)) (kind gifts from Dr. Mikiko Siomi and Dr. Haruhiko Siomi), and rabbit anti-R2D2 (Liu et al. 2003 (link)) (kind gift from Dr. Qinghua Liu), were used as primary antibodies. IRDye 800CW goat anti-mouse IgG, IRDye 800CW goat anti-rat IgG, IRDye 800CW goat anti-rabbit IgG, and IRDye 680RD goat anti-rabbit IgG (Licor) were used as secondary antibodies. The membrane was scanned on an Odyssey imaging system (Licor).

Kandasamy S.K., Zhu L, & Fukunaga R. (2017). The C-terminal dsRNA-binding domain of Drosophila Dicer-2 is crucial for efficient and high-fidelity production of siRNA and loading of siRNA to Argonaute2. RNA, 23(7), 1139-1153.

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Western Blot Analysis of Viral Proteins

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Whole-cell lysates were subjected to SDS–polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes (pore size 0.45 µm, Merck Millipore, MA, USA). PVDF membranes were blocked with 5% skim milk or Blocking One (Nacalai tesque, Kyoto Japan) in Tris-buffered saline containing 0.1% Tween 20 (TBS-T) and probed with anti-Tax mouse monoclonal (Lt-4), anti-HBZ mouse monoclonal (#7-1) [23 (link)], anti-HBZ rat monoclonal (#4B12), anti-β-actin rabbit polyclonal (PM053, MEDICAL & BIOLOGICAL LABORATORIES, Nagoya, Japan), or anti-α-tubulin rabbit polyclonal (PM054, MEDICAL & BIOLOGICAL LABORATORIES) antibodies. PVDF membranes were washed with TBS-T and incubated with IRDye 680RD Goat anti-Mouse IgG, IRDye 800CW goat anti-rat IgG, or IRDye 800CW goat anti-rabbit IgG (LI-COR Biosciences, Lincoln, NE). After washing with TBS-T, the proteins were detected using Odyssey CLx Infrared Imaging System (LI-COR Biosciences).

Naito T., Yasunaga J.I., Mitobe Y., Shirai K., Sejima H., Ushirogawa H., Tanaka Y., Nakamura T., Hanada K., Fujii M., Matsuoka M, & Saito M. (2018). Distinct gene expression signatures induced by viral transactivators of different HTLV-1 subgroups that confer a different risk of HAM/TSP. Retrovirology, 15, 72.

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Phospho-specific GPR35 Antibody Characterization

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A rabbit phospho-site–specific hGPR35 antiserum pSer³⁰⁰/pSer³⁰³-hGPR35a (catalogue number 7TM0102C), raised against the sequence KAHKpSQDpSLCVTL, and a pSer²⁹⁸/pSer³⁰¹-mGPR35 antiserum (7TM0102B), raised against the sequence TPHKpSQDpSQILSLT have been described previously (5 ). A GPR35 (nonphospho) antibody (cat number 7TM0102N), directed against the distal part of the carboxyl-terminal tail of hGPR35 was developed in collaboration with 7TM Antibodies GmbH. IRDye 800CW donkey anti-rabbit immunoglobulin G (IgG), IRDye 800CW donkey anti-goat IgG, and IRDye 800CW goat anti-rat IgG were from LI-COR Biosciences. Horseradish peroxidase anti-mouse (sheep) was from GE Healthcare. High-affinity anti-HA (rat) and anti-HA affinity matrix were from Roche Diagnostics. Anti-LgBiT mAb (cat number N7100) which is an affinity-purified mouse mAb for detection of LgBiT and LgBiT-fusion proteins by Western blotting was purchased from Promega Corporation. GRK isoform–directed antisera for GRK 2 sc-13143 (C-9) c and GRK 5 sc-518005 (D-9) were from Santa Cruz Biotechnology while GRK 3 CS #80362 (D8G6V) and GRK 6 CS #5878 (D1A4) were from Cell Signaling Technology. Further details are found in Reichel et al. (23 (link)).

Ganguly A., Quon T., Jenkins L., Joseph B., Al-awar R., Chevigne A., Tobin A.B., Uehling D.E., Hoffmann C., Drube J, & Milligan G. (2023). G protein–receptor kinases 5/6 are the key regulators of G protein–coupled receptor 35–arrestin interactions. The Journal of Biological Chemistry, 299(10), 105218.

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SDS-PAGE Reagents and Antibodies

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Standard solutions for SDS-PAGE (loading buffer, staining/destaining solutions, running buffers)
Monoclonal ANTI-FLAG M2 antibody produced in mouse (Sigma Aldrich, F3165)
Anti-HA High Affinity from rat IgG1 (Roche, 11867423001)
IRDye 800CW goat anti-rat IgG (LI-COR Biosciences, 925-32219)
IRDye 680RT donkey anti-mouse IgG (LI-COR Biosciences, 925-68072)
Sepax SRT-C 300 column, 5um, 300 A 4.6 × 250 mm (Sepax Technologies, 235300-4625)
N-[4-(7-diethylamino-4-methyl-3-coumarinyl)phenyl]maleimide (CPM) dye (Thermo Fisher, D-10251)

Gustavsson M., Zheng Y, & Handel T.M. (2015). Production of chemokine/chemokine receptor complexes for structural and biophysical studies. Methods in enzymology, 570, 233-260.

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Western Blot Analysis of GIMAP Protein Family

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Cells were lysed in radioimmunoprecipitation assay buffer (150 mM NaCl, 5 mM EDTA [pH 8.0], 50 mM Tris [pH 8.0], 1.0% NP-40, 0.5% sodium deoxycholate, 0.1% SDS) complete with protease and phosphatase inhibitor cocktails, heated at 95°C for 5 min with reducing reagent, and run on a Bis-tris protein gel. Primary antibodies used were anti-β actin (clone 2D1D10; GenScript), anti-HA tag (clone 6E2; Cell Signaling Technologies), anti-LC3 rabbit polyclonal (cat no. L8919; Sigma-Aldrich), anti-human GIMAP1 (PA5-60858; Thermo Fischer Scientific), anti-human GIMAP4 (HPA019137; Sigma-Aldrich), anti-human GIMAP7 (HPA020268; Sigma-Aldrich), and anti-β tubulin Dylight 680 (clone BT7R; Life Technologies). Anti-human and anti-mouse GIMAP6 antibodies (MAC445 and MAC436, respectively), anti-human GIMAP2 antibody, and anti-human GIMAP8 antibody were sourced as previously described (Pascall et al., 2013 (link)). Secondary antibodies used were IRDye 800CW Donkey anti-Mouse (Murine) IgG, IRDye 800CW Donkey anti-Rabbit IgG, and IRDye 800CW Goat anti-Rat IgG (all from Li-COR Biosciences). Membranes were imaged using the Odyssey CLx Imaging System (LI-COR Biosciences) or PXi imager (Syngene). Data were analyzed using Image Studio Lite.

Yao Y., Du Jiang P., Chao B.N., Cagdas D., Kubo S., Balasubramaniyam A., Zhang Y., Shadur B., NaserEddin A., Folio L.R., Schwarz B., Bohrnsen E., Zheng L., Lynberg M., Gottlieb S., Leney-Greene M.A., Park A.Y., Tezcan I., Akdogan A., Gocmen R., Onder S., Rosenberg A., Soilleux E.J., Johnson E., Jackson P.K., Demeter J., Chauvin S.D., Paul F., Selbach M., Bulut H., Clatworthy M.R., Tuong Z.K., Zhang H., Stewart B.J., Bosio C.M., Stepensky P., Clare S., Ganesan S., Pascall J.C., Daumke O., Butcher G.W., McMichael A.J., Simon A.K, & Lenardo M.J. (2022). GIMAP6 regulates autophagy, immune competence, and inflammation in mice and humans. The Journal of Experimental Medicine, 219(6), e20201405.

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Immuno-blotting of protein samples

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Protein concentration was measured using a Direct Detect Infrared Spectrometer (Merck). 20 µg of proteins were separated on a NuPAGE 4–12% Bis-Tris gel (Thermo Fisher Scientific). Proteins were transferred with an iBLot2 device (Invitrogen) on a nitrocellulose membrane and blocked for 1 hr in 1x Licor TBS Blocking Buffer (Licor). Primary antibodies were incubated over night at 4°C. Licor secondary antibodies were incubated for 45 min at room temperature (RT) and images acquired with an Odyssey CLx scanner (Licor). The following antibodies were used: anti-HA (ab9110), anti-FLAG (Sigma #F1804), anti-GFP (ab13970), anti-Piwi (described in Brennecke et al., 2007 (link)), anti-Nxt1 (Herold et al., 2001 (link)), anti-Histone H3 (ab10799), anti-Tubulin (ab18251), mouse anti-Panx (Sienski et al., 2015 (link)), IRDye 680RD Donkey anti-Rabbit IgG (H + L) (Licor), IRDye 800CW Donkey anti-Mouse IgG (H + L) (Licor), IRDye 800CW Goat anti-Rat IgG (H + L) (Licor).

Fabry M.H., Ciabrelli F., Munafò M., Eastwood E.L., Kneuss E., Falciatori I., Falconio F.A., Hannon G.J, & Czech B. (2019). piRNA-guided co-transcriptional silencing coopts nuclear export factors. eLife, 8, e47999.

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Antibody Characterization for FAM83 and CK1

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Antibodies recognising FAM83B (SA270), FAM83D (SA102), FAM83F (SA103), FAM83H (SA273), CK1α (SA527), CK1ε (SA610), CK1δ (SA609), and GFP (S268B) were generated in-house and are available for request from the MRC-PPU reagents website (http://mrcppureagents.dundee.ac.uk). Antibodies recognising GAPDH (14C10) (#2118), IKZF1 (D6N9Y) (#14859), CRBN (D8H3S) (#71810), Na, K-ATPase alpha1 (D4Y7E) (#23565), and Lamin A/C (#2032) were obtained from Cell Signalling Technology. Additional antibodies used were FAM83G (ab121750; Abcam), α-tubulin (MA1-80189; Thermo Fisher Scientific), Ubiquitin (BML-PW8810; Enzo), HIF-1α (6109590; BD Biosciences), and ZFP91 (A303-245A; Bethyl Laboratories). Secondary antibodies used were StarBright Blue 700 goat anti-rabbit IgG (12004161; Bio-Rad), StarBright Blue 700 goat anti-mouse IgG (12004158; Bio-Rad), IRDye 800CW donkey anti-goat IgG (926-32214; LI-COR) and IRDye 800CW goat anti-rat IgG (926-32219; LI-COR).

Dunbar K., Macartney T.J, & Sapkota G.P. (2020). IMiDs induce FAM83F degradation via an interaction with CK1α to attenuate Wnt signalling. Life Science Alliance, 4(2), e202000804.

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Antibodies for Cellular and Molecular Analysis

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Antibodies recognising GFP (S268B), CK1α (SA527), CK1δ (SA609), CK1ε (SA610), and FAM83F (SA103) are available on request from the MRC-PPU reagents website (http://mrcppureagents.dundee.ac.uk). Antibodies for β-actin (#4967), GAPDH (14C10) (#2118), p-LRP6 (S1490) (#2568), LRP6 (C47E12) (#3395), β-catenin (D10A8) (#8480), phospho-β-catenin (S45) (#9564), phospho-β-catenin (S33/S37/T41) (#9561), Na/K ATPase (D4Y7E) (#23565), and Lamin A/C (#2032) were purchased from Cell Signalling Technology. Additional antibodies used were FAM83G (ab121750; Abcam), α-tubulin (MA1-80189; Thermo Fisher Scientific), Active-β-catenin (anti-ABC) clone 8E7 (05-665; END Millipore), mouse anti-GFP clone 7.1 and 13.1 (Roche), mouse-monoclonal anti-HA (H9658; Sigma-Aldrich), and U2AF1 (PA5-28510; Thermo Fisher Scientific). Secondary antibodies used were StarBright Blue 700 Goat anti-Rabbit IgG (12004161; Bio-Rad), StarBright Blue 700 Goat anti-Mouse IgG (12004158; Bio-Rad), IRDye 800CW Donkey anti-Goat IgG (926-32214; LI-COR), IRDye 800CW Goat anti-Rat IgG (926-32219; LI-COR), IRDye 800CW Goat anti-Mouse (926-32210; LI-COR), and IRDye 680LT Goat anti-Rabbit (926-68021; LI-COR).

Dunbar K., Jones R.A., Dingwell K., Macartney T.J., Smith J.C, & Sapkota G.P. (2020). FAM83F regulates canonical Wnt signalling through an interaction with CK1α. Life Science Alliance, 4(2), e202000805.

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