Peroxidase conjugated affinipure goat anti rabbit igg

Manufactured by Jackson ImmunoResearch

Sourced in United States

Peroxidase-conjugated AffiniPure goat anti-rabbit IgG is a secondary antibody product that binds to rabbit immunoglobulin G (IgG) and is conjugated with the enzyme peroxidase. This product can be used in various immunoassay and detection techniques that involve the use of rabbit primary antibodies.

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Lab products found in correlation

Peroxidase conjugated affinipure goat anti mouse igg, by Jackson ImmunoResearch (5 mentions) Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions) Skim milk, by Cell Signaling Technology (1 mentions) Anti gapdh antibody, by Cell Signaling Technology (1 mentions) Skim milk, by BD (1 mentions) Anti smad4, by Cell Signaling Technology (1 mentions) Costar eia ria plates, by Corning (1 mentions) Carbonate bicarbonate buffer, by Merck Group (1 mentions) 1 step ultra tmb elisa solution, by Thermo Fisher Scientific (1 mentions) Alexa flour 488 conjugated, by Thermo Fisher Scientific (1 mentions) Anti flag mab f3165, by Merck Group (1 mentions) Anti flag pab f7425, by Merck Group (1 mentions) Anti gfp pab, by Thermo Fisher Scientific (1 mentions) Fluorescein isothiocyanate conjugated affinipure goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Lipofectamine 2000, by Wisent (1 mentions) Cofilin, by Cell Signaling Technology (1 mentions) Tsg101, by Thermo Fisher Scientific (1 mentions) Hsc70, by Enzo Life Sciences (1 mentions) Flag tag (flag), by Merck Group (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions)

12 protocols using peroxidase conjugated affinipure goat anti rabbit igg

Silver Nanoparticles Induce Stress Responses

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Cells were treated with 5 nm and 100 nm silver NPs for 24 h. Cells were harvested and lysed using lysis buffer (200 µL; 150 mM NaCl, 1% NP-40, 0.1% sodium dodecyl sulfate (SDS), 50 mM Tris (pH 8), 5 mM NaF, 1 mM Na₃VO₄, 1 mM PMSF, protease inhibitor cocktail) at 4 °C for 2 h. Cell lysates were centrifuged at 13,000 rpm at 4 °C for 15 min, and the supernatants were stored at −20 °C. Protein (30–40 µg) from each sample was boiled for 5 min and loaded on a 12% SDS–polyacrylamide gel. After electrophoresis, proteins in the gel were transferred onto nitrocellulose membranes (Amersham, Glattbrugg, Switzerland). After blocking with 5% skim milk (BD Biosciences, Franklin, NJ, USA), membranes were reacted overnight with primary antibodies, including anti-HO-1 antibodies (Cell Signaling Technology, Danvers, MA, USA) and anti-HSP-70 antibodies (Cell Signaling Technology), at 1:1000 dilution in 5% bovine serum albumin (Affymetrix, Cleveland, OH, USA) at 4 °C. After washing, the membranes were further reacted with a secondary antibody (peroxidase-conjugated AffiniPure goat anti-rabbit IgG; Jackson ImmunoResearch, West Grove, PA, USA) at 1:2000 dilution in 5% skim milk at room temperature for 1 h. Anti-GAPDH antibody (Cell Signaling Technology) was used to assess the transcription levels of housekeeping genes.

Jang J., Park S, & Choi I.H. (2021). Increased Interleukin-11 and Stress-Related Gene Expression in Human Endothelial and Bronchial Epithelial Cells Exposed to Silver Nanoparticles. Biomolecules, 11(2), 234.

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Western Blot Analysis of Smad4 and Akirin1

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The primary antibodies used for Western blotting were (1) rabbit polyclonal anti-Smad4 (1 : 1000; cat# 9515; Cell Signaling Technology), (2) rabbit polyclonal anti-akirin1 (1 : 1000; cat# ab77075; Abcam) and (3) rabbit polyclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1 : 2000; cat# ABS16; Millipore, Temecula, CA, USA). The corresponding secondary antibody used for all Western blots was peroxidase-conjugated AffiniPure goat anti-rabbit IgG (1 : 10 000; cat# 111-035-003; Jackson Immuno Research Laboratories).

Silva M.T., Wensing L.A., Brum P.C., Câmara N.O, & Miyabara E.H. (2014). Impaired structural and functional regeneration of skeletal muscles from β2-adrenoceptor knockout mice. Acta Physiologica (Oxford, England), 211(4), 617-633.

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Quantifying hrRNASET2 Binding to Actin

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hrRNASET2 binding to actin was quantified as modified from Liu et al. [22 (link)]. Human platelet actin (5 ug/ml) (Cytoskeleton, Denver, CO, USA) was diluted in carbonate-bicarbonate buffer (Sigma-Aldrich, St Louis, MO, USA), pH 9.5, and coated directly onto 96-wells COSTAR EIA/RIA plates (Corning, NY, USA) overnight at 4°C. The plates were then blocked with 5% (w/v) BSA in PBS containing 0.25% Tween-20 (PBST), at room temperature for 1 h, and subsequently incubated with hrRNASET2, at 1:2 serial dilutions in PBST, overnight at 4°C. Plates were washed three times with TBST, under continuous shaking for 10 min each, at room temperature and then incubated with rabbit anti-RNASET2 polyclonal affinity pure antibodies (GENEMED SYNTHESIS, San Antonio, TX, USA), diluted 1:500 in PBST at 37°C for 1 h. After three washing as described above, 0.8 μg/ml peroxidase-conjugated AffiniPure goat anti-rabbit IgG (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) in TBST was added and incubated under the same conditions. Plates were then washed twice with TBST and once with TBS, as described above. 1-Step Ultra TMB-ELISA solution (Thermo Scientific, Pierce Biotechnology, Rockford, IL, USA) was then added and optical absorbance was detected at 650 nm with an Infinite F50 multidetection microplate reader (Tecan, Grödig, Austria).

Roiz L., Smirnoff P., Lewin I., Shoseyov O, & Schwartz B. (2016). Human recombinant RNASET2: A potential anti-cancer drug. Oncoscience, 3(2), 71-84.

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Characterization of nApoECFp17 Antibody

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The in house, rabbit polyclonal nApoECFp17 antibody was synthesized and extensively characterized as previously described and detects the amino-terminal fragment of APOE corresponding to amino acids 1-151 [11 (link)]. The nApoECFp17 antibody was synthesized using the 7-mer peptide C-RKRLLRD, which represents the N-terminal upstream neoepitope fragment of APOE4 and E3 that would be generated following cleavage after the terminal aspartic acid residue at position D151. The anti-APOE4 N-terminal rabbit polyclonal antibody was purchased from Aviva Systems Biology Corp. (San Diego, CA). Peroxidase-conjugated AffiniPure goat anti-rabbit (IgG) was purchased from Jackson ImmunoResearch Labs (West Grove, PA). This antibody is known to recognize the extreme N-terminal region of human APOE. The TMB microwell peroxidase substrate system was purchased from KPL, Inc. (Gaithersburg, MD). 96-well, clear, flat bottom, high binding, polystyrene plates were purchased from Corning (Kennebunk, ME).

Gause J., Day R., Caraway C., Poon W, & Rohn T. (2017). Evaluation of Apolipoprotein E Fragmentation as a Biomarker for Alzheimer’s Disease. Journal of neurology and neurological disorders, 3(2), 204.

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Anti-PRRSV nsp1β Rabbit Antibody Generation

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An anti-PRRSV-nsp1β rabbit pAb specific for NA-PRRSV nsp1β was generated at the Immunological Research Center, University of Illinois at Urbana-Champaign (Urbana, IL). The anti-LV-nsp2/3 antibody was a generous gift from E. Snijder, (Leiden University Medical Center, Leiden, Netherlands). Anti-N protein mAbs (SDOW17) was obtained from E. Nelson (South Dakota State University, Brookings, SD). polyinosinic: polycytidylic [poly (I:C)], DAPI (4′,6-diamidino-2-phenylindol), anti-Flag mAb (F3165), and anti-FLAG pAb (F7425) were purchased from Sigma (St. Louis, MO). Anti-β-actin mAb (sc-47778), anti- HSP90 mAb (sc-69703), and anti-PARP pAb (sc-7150) were purchased from Santa Cruz Biotechnologies Inc. (Santa Cruz, CA). Anti-ISG15 pAb and anti-GFP pAb were purchased from Thermo Scientific (Rockford, IL). Anti-DEPTOR pAb was purchased from Invitrogen (Carlsbad, CA). The peroxidase-conjugated Affinipure goat anti-mouse IgG and peroxidase-conjugated Affinipure goat anti-rabbit IgG were purchased from Jackson Immuno Research (West Grove, PA). Alexa-Flour 488-conjugated and Alexa-Flour 568-conjugated secondary antibodies were purchased from Invitrogen.

Han M., Ke H., Zhang Q, & Yoo D. (2017). Nuclear imprisonment of host cellular mRNA by nsp1β protein of porcine reproductive and respiratory syndrome virus. Virology, 505, 42-55.

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Antibody Reagents for Cell Adhesion Studies

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Mouse anti‐vinculin (clone V9131), rabbit anti‐paxillin (SAB4502553), rabbit anti‐integrin α3 (AB1920), and rabbit anti‐actin IgGs (A2066) were purchased from Millipore‐Sigma (Oakville, ON). Mouse anti‐human talin‐1 IgG (clone 97H6) was purchased from Bio‐Rad Laboratories (Mississauga, ON). Rabbit monoclonal antibody to phospho‐FAK Y397 (31H5L17) was purchased from ThermoFisher Scientific (Saint‐Laurent, QC). Rat anti‐collagen IV α5 clone H53 (7078) was purchased from Chondrex Inc. (Woodinville, WA). Rabbit anti‐nephrin antiserum was kindly provided by Dr. Tomoko Takano (McGill University). Fluorescein isothiocyanate‐conjugated AffiniPure goat anti‐rabbit IgG (#111‐095‐003), rhodamine‐conjugated AffiniPure goat anti‐mouse IgG (#415‐505‐1), peroxidase‐conjugated AffiniPure sheep‐anti mouse IgG (#515‐035‐003), and peroxidase‐conjugated AffiniPure goat anti‐rabbit IgG (#111‐035‐003) were purchased from Jackson Immunoresearch (West Grove, PA).
Electrophoresis reagents were from Bio‐Rad Laboratories (Mississauga, ON), and GE Healthcare (Baie d'Urfé, QC). Tissue culture media and Lipofectamine 2000 were from Wisent (Saint‐Jean‐Baptiste, QC) and Invitrogen‐Life Technologies (Burlington, ON).

Cybulsky A.V., Papillon J., Bryan C., Navarro‐Betancourt J.R, & Sabourin L.A. (2024). Role of the Ste20‐like kinase SLK in podocyte adhesion. Physiological Reports, 12(1), e15897.

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Antibody Sourcing and Validation Protocol

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Antibodies were purchased from the following commercial sources: Aβ (Cell Signaling Technology, 8243S), Tau (Santa Cruz Biotech, sc-390476), Hsc70 (Enzo Life Sciences, ADI-SPA-815-F), TSG101 (Thermo Fisher, PA5-31260), GFP (Cell Signaling Technology, 2956S), β-Actin (Santa Cruz Biotech, sc-47778), p62 (Cell Signaling Technology, 5114), Flag (Sigma Aldrich, F1804), SSH1 (ECM Biosciences, SP1711) (Cell Signaling Technology, 13578), Cofilin (Cell Signaling Technology, 5175s), Peroxidase-Conjugated AffiniPure Goat anti-mouse IgG (Jackson ImmunoResearch, 115-035-033), Peroxidase-conjugated AffiniPure Goat anti-rabbit IgG (Jackson ImmunoResearch, 111-035-033), donkey anti-rat IgG-HRP (Southern Biotech, 6430-05).

Cazzaro S., Fang C., Khan H., Witas R., Kee T.R., Woo J.A, & Kang D.E. (2022). Slingshot homolog-1 mediates the secretion of small extracellular vesicles containing misfolded proteins by regulating autophagy cargo receptors and actin dynamics. Frontiers in Aging Neuroscience, 14, 933979.

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Immunoblotting of LHCSR3 and HA-tagged Proteins

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Cells were harvested by centrifugation at 12,000 × g for 30 s, and resuspended in 60 μl of SBA buffer (100 mM DTT, 100 mM Na₂CO₃), with 40 μl of SBB buffer (30% sucrose, 5% SDS). The samples were vortex for 20 min at room temperature and then subjected to 3 freeze/thaw cycles. After centrifugation, the supernatants were loaded on a 10%−12.5% SDS-PAGE gel and the proteins were blotted onto a nitrocellulose membrane. Membranes were blocked for 0.5 h with 5% milk in TBST and then incubated with anti-LHCSR3 polyclonal antibody (Agrisera), diluted 1:10,000 in TBST or anti-HA mono-clonal antibody (Cell Signaling Technology), diluted 1:1,000, anti-α-Tubulin mono-clonal antibody (Sigma) diluted 1:1,000 for one hour and then rinsed three times for 5 min before incubation with peroxidase-conjugated affinipure goat anti-rabbit IgG (Jackson) or peroxidase-conjugated affinipure goat anti-mouse IgG (Jackson) both diluted 1:10,000 for 1 hour. The blots were developed with ECL detection reagent (Millipore) and images of the blots were obtained using a CCD imager (Thermo).

Xue J.H., Chen G.D., Hao F., Chen H., Fang Z., Chen F.F., Pang B., Yang Q.L., Wei X., Fan Q.Q., Xin C., Zhao J., Deng X., Wang B.A., Zhang X.J., Chu Y., Tang H., Yin H., Ma W., Chen L., Ding J., Weinhold E., Kohli R.M., Liu W., Zhu Z.J., Huang K., Tang H, & Xu G.L. (2019). A vitamin C-derived DNA modification catalyzed by an algal TET homolog. Nature, 569(7757), 581-585.

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Cardiac Protein Expression Analysis

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Primary antibodies (1/2000) included monoclonal mouse anti-SERCA2 ATPase (2A7-A1; MA3-919), monoclonal mouse anti-Phospholamban (2D12; MA3-922), polyclonal rabbit anti-Calsequestrin (PA1-913) and monoclonal mouse anti-Na⁺,K⁺-ATPase alpha-3 (XVIF9-G10; MA3-915) from Thermo scientific, monoclonal mouse anti-Kir2.2 (S24-1; ab84821) from Abcam, monoclonal mouse anti-Kir2.3 (N25/35; 75-069) from Neuromab, and monoclonal anti-GAPDH (10R-G109a) from Fitzgerald. Peroxidase-conjugated AffiniPure goat anti-rabbit IgG (111-035-144) and Affinipure donkey anti-mouse IgG (715-035-151) from Jackson ImmunoResearch were used as secondary antibodies (1/10,000).

Sivagangabalan G., Nazzari H., Bignolais O., Maguy A., Naud P., Farid T., Massé S., Gaborit N., Varro A., Nair K., Backx P., Vigmond E., Nattel S., Demolombe S, & Nanthakumar K. (2014). Regional Ion Channel Gene Expression Heterogeneity and Ventricular Fibrillation Dynamics in Human Hearts. PLoS ONE, 9(1), e82179.

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Antibody-Based Investigation of PRRSV Signaling

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The antibodies and chemicals used in the present study are listed as follow. Anti (α)-PRRSV-N MAb (MR40) (mouse) was obtained from E. Nelson (South Dakota State University, Brookings, SD). α-p65 PAb (rabbit) (H-286, sc-7151), α-p65 MAb (mouse) (F-6, sc-8008), α-PIAS1 MAb (mouse) (F-1, sc-365127), α-PARP PAb (rabbit) (H-250, sc-7150), α-HSP90 MAb (mouse) (4F10, sc-69703), α-SUMO1 MAb (mouse) (D-11, sc-5308), α-SUMO2/3 PAb (rabbit) (FL-103, sc-32873), and α-β-actin MAb (mouse) (C4, sc-47778) were purchased from Santa Cruz Biotechnologies Inc. (Santa Cruz, CA). α-FLAG MAb (rat) (M2) was purchased from Agilent Technologies Inc. (Cedar Creek, TX). α-FLAG PAb (rabbit) was purchased from Rockland Inc. (Gilbertsville, PA). α-GFP MAb (rabbit), α-HA MAb (mouse), α-PIAS1 PAb (rabbit), and Alexa-Flour 488-conjugated and Alexa-Flour 568-conjugated secondary antibodies were purchased from ThermoFisher (Rockford, IL). α-HA MAb (rabbit) (C29F4), α-phospho-NF-κB p65 (Ser 536) MAb (rabbit) (93H1), and human Tumor Necrosis Factor-α (TNF-α) (8902) were purchased from Cell Signaling (Danvers, MA). The peroxidase-conjugated Affinipure goat anti-mouse IgG and peroxidase-conjugated Affinipure goat anti-rabbit IgG were purchased from Jackson Immuno Research (West Grove, PA). Lipopolysaccharides (LPS) and DAPI (4′, 6′-diamidino-2-phenylindol) were purchased from Sigma (St. Louis, MO).

Ke H., Lee S., Kim J., Liu H.C, & Yoo D. (2019). Interaction of PIAS1 with PRRS virus nucleocapsid protein mediates NF-κB activation and triggers proinflammatory mediators during viral infection. Scientific Reports, 9, 11042.

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