Biotin conjugated goat anti rabbit igg

Manufactured by Thermo Fisher Scientific

Sourced in United States

Biotin-conjugated goat anti-rabbit IgG is an antibody reagent used for immunodetection and immunoassay applications. It is produced by conjugating biotin to goat-derived antibodies that are specific for rabbit immunoglobulin G (IgG).

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6 protocols using biotin conjugated goat anti rabbit igg

Immunohistochemical Analysis of Oxidative Stress

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The immunohistochemistry staining methods were performed as previously described35 (link). After dewaxing, antigen retrieval and blocking of endogenous peroxidase, the sections were fixed with goat serum and incubated with a 1:1000 dilution of rabbit monoclonal anti-CGL antibody (Abcam, Cambridge MA, USA), a 1:2000 dilution of rabbit monoclonal anti-CBS antibody (Abcam, Cambridge MA, USA), and a 1:20 dilution of mouse monoclonal anti-8-OHdG antibody (Santa cruz biotechnology, CA, USA) overnight at 4 °C, followed by incubation with biotin-conjugated goat anti-rabbit IgG (Invitrogen, Carlsbad, CA, USA) and streptavidin- conjugated peroxidase (Invitrogen, Carlsbad, CA, USA). Finally, sections were detected under a microscope after reaction using 3,3′-diaminobenzidine (DAB) (Invitrogen, Carlsbad, CA, USA). The intensity of immunohistochemical staining of 8-OHdG was analyzed in ten random fields (×400) of renal cortex per rat using Image J software (NIH, MD, USA).

Wang W.J., Cai G.Y., Ning Y.C., Cui J., Hong Q., Bai X.Y., Xu X.M., Bu R., Sun X.F, & Chen X.M. (2016). Hydrogen sulfide mediates the protection of dietary restriction against renal senescence in aged F344 rats. Scientific Reports, 6, 30292.

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Immunolabeling of Adult Fly Brain

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An adult fly brain was prepared and imaged as previously described^{21 (link)}. The dissected brain was incubated with primary antibody at 4 °C for two days, and the dilution ratios were 1:10 for mouse 4F3 anti-DLG (Hybridoma Bank, University of Iowa), 1:500 for rabbit anti-HA (Abcam), and 1:500 for rabbit anti-GFP (Invitrogen). The secondary antibody solution with the fly brain was incubated at room temperature overnight, and the dilution ratio was 1:200 for biotin-conjugated goat anti-mouse IgG (Invitrogen) and biotin-conjugated goat anti-rabbit IgG (Invitrogen), and then the fly brain was labeled by 1:200 dilution with Alexa Fluor 635 streptavidin (Invitrogen) at room temperature overnight.

Wu J.K., Tai C.Y., Feng K.L., Chen S.L., Chen C.C, & Chiang A.S. (2017). Long-term memory requires sequential protein synthesis in three subsets of mushroom body output neurons in Drosophila. Scientific Reports, 7, 7112.

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Recombinant CCN1 Proteins Production

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Recombinant CCN1 and mutant proteins (D125A, DM, CCN1-I.II, CCN1-ΔCT, and D125A-ΔCT) were produced using a baculovirus expression system in insect cells and purified by ion-exchange or immuno-affinity chromatography^{30 (link), 32 (link)}. Rabbit polyclonal antibodies against CCN1 were raised against the CCN1 vWC domain^{67 (link)}. Rabbit and mouse IgGs, echstatin, and lipopolysacharide (LPS) were from Sigma. RGDSP and RGESP peptides were from Bachem. Function-blocking mAbs against mouse integrin β₁ (KM16), α_v (RMV-7), β₃ (2C9-G3), α_vβ₃ (23-C6) and β₅ (KN52) were from eBioscience. Recombinant murine interleukin-4 (IL-4) were from R&D systems. Horse radish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit secondary antibodies were from Amersham Biosciences. Biotin-conjugated goat anti-rabbit IgG, biotin-conjugated goat anti-rat IgG antibody, and streptavidin-conjugated HRP were from Invitrogen.

Jun J.I., Kim K.H, & Lau L.F. (2015). The Matricellular Protein CCN1 Mediates Neutrophil Efferocytosis in Cutaneous Wound Healing. Nature communications, 6, 7386.

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Recombinant CCN1 Proteins Production

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Jun J.I., Kim K.H, & Lau L.F. (2015). The Matricellular Protein CCN1 Mediates Neutrophil Efferocytosis in Cutaneous Wound Healing. Nature communications, 6, 7386.

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Immunohistochemistry and Immunofluorescence Protocols

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Cell culture media- RPMI 1640 (Thermo Fisher Scientific, Gibco, NY, 11875093), DPBS, 0.25% (w/v) Trypsin, (100X), and Penicillin-Streptomycin (5,000 U/ml) were purchased from ThermoFisher Scientific. Fetal bovine serum was purchased from GIBCO. For immunohistochemistry, biotin-conjugated goat anti-rabbit IgG (Invitrogen, 31820) was used as a secondary antibody followed by Reagent A: Avidin (Thermo Scientific, 1852280) and Reagent B: Biotinylated HRP (Thermo Scientific, 1852310). ImPACT DAB (Vector Laboratories, SK-4105) was used for visualizing the protein. For immunofluorescence and immunocytochemistry, secondary Alexa Fluor 594 goat anti-mouse (Thermo Fisher A11020), and Alexa Fluor 488 goat anti-rabbit (Thermo Fisher A11008) antibodies were used. Other reagents such as HEPES, KCl, DTT, NP-40, Glycerol, MgCl2, EDTA, PMSF, protease inhibitors such as aprotinin, and leupeptin were purchased from Sigma-Aldrich. Halt phosphatase inhibitor (1862495), Trizol and Powerup SYBR Green master mix were purchased from ThermoFisher Scientific. cDNA kit was purchased from Roche. All the primers were from IDT.

Dutta S., Bhattacharya S., Harris H., Islam R., Bodas S., Polavaram N., Mishra J., Das D., Seshacharyulu P., Kalluchi A., Pal A., Kohli M., Lele S., Muders M., Batra S., Ghosh P., Datta K, & Rowley M. (2023). Understanding the role of Pax5 in development of taxane-resistant neuroendocrine like prostate cancers. Research Square.

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Vascular Calcification Analysis in Mice

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Innominate arteries from 12 week old ApoE−/−OPG−/− and ApoE−/−OPG+/+ mice were fixed in formalin and embedded in paraffin. Five micron thick sections were used for histochemical and immunohistochemical analyses. Alizarin Red S or Von Kossa staining was used to detect calcium deposition. The following antibodies were employed: anti goat SMα-actin (Sigma, St Louis, MO), goat anti SM22α (Abcam, Cambridge, MA), rabbit anti phosphorylated ERK (CellSignaling, Danvers, MA), goat anti Runx2 (R&D Systems, Minneapolis, MN), mouse anti RANKL (Imgenex, San Diego, CA), goat anti Osteopontin (R&D Systems, Minneapolis, MN), rabbit anti cleaved caspase-3 (CellSignaling, Danvers, MA), control rabbit, goat and mouse IgG (Invitrogen, Carlsbad, CA), biotin conjugated rabbit anti-goat IgG (Pierce Thermoscientific, Rockford, IL), biotin conjugated goat anti-rabbit IgG (Invitrogen, Carlsbad, CA), biotin conjugated rabbit anti-mouse IgG (Invitrogen, Carlsbad, CA). Sections were counterstained with haematoxylin (Ricca Chemical Company, Arlington, TX) or Methyl Green (Sigma).

Callegari A., Coons M.L., Ricks J.L., Rosenfeld M.E, & Scatena M. (2014). Increased Calcification in Osteoprotegerin Deficient Smooth Muscle Cells: Dependence on Receptor Activator of NF-κB Ligand and Interleukin-6. Journal of vascular research, 51(2), 118-131.

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