Goat anti rabbit 111 035 003

Manufactured by Jackson ImmunoResearch

Sourced in United States

Goat anti-rabbit (111-035-003) is a secondary antibody product produced by Jackson ImmunoResearch. It is a polyclonal antibody that recognizes rabbit immunoglobulins and is conjugated with horseradish peroxidase (HRP) for use in various immunoassay applications.

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Close spelling variants of this product

Goat Anti-Rabbit IgG (111-035-003) (3 citations) Peroxidase-AffiniPure goat anti-rabbit IgG (111-035-003), (2 citations) Peroxidase AffiniPure Goat Anti-Rabbit IgG(H+L) 111-035-003 (2 citations) Goat anti-mouse (115-035-003) and goat anti-rabbit (111-035-003) secondary antibodies (2 citations) Anti-rabbit (111-035-003, (5 citations)

8 protocols using goat anti rabbit 111 035 003

Molecular Mechanisms of JAK2-FOXO3 Pathway

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Mouse anti-β-actin, mouse anti-Flag, and mouse anti-HA antibody and the following chemicals and solvents (MG132, cycloheximide, dimethyl sulfoxide (DMSO), glycerol, glycine, sodium chloride, Trizma base, and Tween20) were from Sigma (St. Louis, MO, USA). AZD1480 was from Selleckchem (Houston, TX, USA). Rabbit anti-IL4Rα, rabbit anti-IL13Rα1, mouse anti-PARP1, rabbit anti-FOXO3, mouse anti-Lamin B1, and mouse anti-GAPDH antibodies were from Santa Cruz Biotechnology. Rabbit anti-JAK2, rabbit anti-pJAK2, rabbit anti-Tyr, rabbit anti-cleaved PARP1, rabbit anti-cleaved Caspase3, rabbit anti-Bax, rabbit anti-Bim, rabbit anti-Bcl2, rabbit anti-p21, and rabbit anti-p27 antibodies were from Cell Signaling (Danvers, MA, USA). Goat anti-rabbit (111-035-003) and goat anti-mouse (115-035-003) horseradish peroxidase-conjugated IgG were obtained from Jackson ImmunoResearch (West Grove, PA, USA). Enhanced chemiluminescence (ECL) reagents were obtained from Genedepot (Barker, TX, USA). pECE empty/Flag-FOXO3 and pCMV3-C-HA empty/HA-JAK2 plasmid DNA were from Addgene (Watertown, MA, USA) and Sino Biological (Wayne, PA, USA), respectively.

Kang M.A., Lee J., Ha S.H., Lee C.M., Kim K.M., Jang K.Y, & Park S.H. (2019). Interleukin4Rα (IL4Rα) and IL13Rα1 Are Associated with the Progress of Renal Cell Carcinoma through Janus Kinase 2 (JAK2)/Forkhead Box O3 (FOXO3) Pathways. Cancers, 11(9), 1394.

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Western Blot Analysis of Apoptosis Markers

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Cells or tumor tissue samples were lysed with RIPA buffer (Beyotime) containing protease inhibitors cocktail (Selleck). After centrifugation, samples were loaded on to and separated using SDS/PAGE, then transferred on to PVDF membranes (Millipore). The membranes were blocked for 1 h at room temperature and incubated with the anti-β-actin (#3700, Cell Signaling Technology), anti-PARP1 (13371-1-AP, Proteintech) and anti-Caspase 3 (#9662, Cell Signaling Technology) antibodies overnight at 4°C. After washing, the blots were incubated with goat anti-rabbit (111-035-003, Jackson) or anti-mouse (115-035-003, Jackson) HRP-conjugated secondary antibodies and visualized using the Immobilon™ HRP Substrate Peroxide Solution (Millipore).

Qi H., Lu Y., Lv J., Wu H., Lu J., Zhang C., Zhang S., Bao Q., Zhang X., Xie C, & Yin Z. (2018). The long noncoding RNA lncPARP1 contributes to progression of hepatocellular carcinoma through up-regulation of PARP1. Bioscience Reports, 38(3), BSR20180703.

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Western Blot Analysis of Keap1, Nrf2, and HO-1

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HaCaT and PIG1 cells were lysed in RIPA lysis buffer on ice. Equal amounts of protein from each sample were separated using 10% SDS-PAGE and then transferred to polyvinylidene fluoride membranes (Merck Millipore, Billerica, MA, USA). In addition, 5% bovine serum albumin (BSA) (Yeasen, Shanghai, China) was used to block nonspecific binding for 1 h. The membranes were probed with anti-Keap1 (ab227828), anti-Nrf2 (ab62352), anti-HO-1 (ab13248), and anti-β-actin (ab8227) antibodies overnight at 4°C; these primary antibodies were obtained from Abcam (Cambridge, MA, USA). Membranes were then incubated with secondary antibodies, including goat anti-mouse (115-035-003; Jackson ImmunoResearch, PA, USA) and goat anti-rabbit (111-035-003, Jackson ImmunoResearch, PA, USA) antibodies, at room temperature for 1 h. Thereafter, the protein bands were examined using an ECL western blot detection system (Merck Millipore, Burlington, MA, USA). The antibodies used are listed in Supplementary Table 2.

Xie Y., Chen Z, & Wu Z. (2022). Four-Octyl Itaconate Attenuates UVB-Induced Melanocytes and Keratinocytes Apoptosis by Nrf2 Activation-Dependent ROS Inhibition. Oxidative Medicine and Cellular Longevity, 2022, 9897442.

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Quantifying Autophagy Pathway Activation

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Chloroquine (C6628) was obtained from Sigma-Aldrich. Oxaliplatin (OXA, S1224), and quinacrine (S5435) were purchased from Selleckchem. Torin 1 (2273-5) was ordered from BioVision. Anti-ATF4 (CST 11815S), anti-CHOP antibody (2895), anti-eIF2α antibody (9722), anti-p-eIF2α (Ser51) (9721S), anti-Histone H3 (9715S), anti-ATG5 antibody (12994), and anti-PERK (5683) antibodies were obtained from Cell Signaling Technology (CST). An anti-LC3 antibody (NB100-2220) was obtained from Novus Biologicals. An anti-GAPDH antibody (GTX100118) was ordered from GeneTex. Anti-TFE3 antibody (HPA023881), and anti-Flag (F3165) antibody were ordered from Sigma-Aldrich. Anti-TFEB antibody (303-673A) was ordered from Bethyl Laboratories. Anti-HMGB1 (ab18256), and anti-Calreticulin (ab92516) antibodies were ordered from Abcam. Goat anti-rabbit (111-035-003), and goat anti-mouse (115-035-003) secondary antibodies were products of Jackson ImmunoResearch. Alexa Fluor 594 goat anti-rabbit IgG (A-11012), Alexa Fluor 488 goat anti-rabbit IgG (A-11034), and Alexa Fluor 488 (A-11008) goat anti-rabbit secondary antibodies were ordered from Invitrogen.

Wang S., Wang G., Wu W., Xu Z., Yang J., Cao M., Wang Q., Wang J., Yang C, & Zhang W. (2022). Autophagy activation by dietary piceatannol enhances the efficacy of immunogenic chemotherapy. Frontiers in Immunology, 13, 968686.

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Biochemical Reagents for Cell Signaling

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Biochemical reagents, such as Cytochalasin B (C-6762), 2-deoxyglucose (D-8375), Dexamethasone (D2915), isobutymethylxanthine (I5879), Insulin (I0516), Rosiglitazone (R2408), BCS (Bovine Calf serum, 12133C), BSA (Bovine Serum Albumin, A9647) and Oil Red O (O0625), were all purchased from Sigma-Aldrich (St. Louis, MO, USA). 2-[3H] deoxyglucose was purchased from Perkin Elmer (NET328250UC). All lentiviral plasmids harboring shRNA sequences (mission shRNA) were obtained from Sigma. Antibodies against β-actin (Sigma-Aldrich, St Louis, MO, USA; a5441), α-tubulin (clone B-5-1-2) and total PKN1 were purchased from Sigma-Aldrich. Phospho-IRS-1 (Tyr895), Phospho-PRK1 (Thr774)/PRK2 (Thr816), Phospho-Akt (Ser473) and Phospho-p70 S6 kinase (Thr 389) were purchased from cell signaling. Peroxidase conjugated goat anti-mouse (115-035-003) and goat anti-rabbit (111-035-003) antibodies were obtained from Jackson Immunoresearch (West Grove, PA, USA).

Herrerías-González F., Yeramian A., Baena-Fustegueras J.A., Bueno M., Fleitas C., de la Fuente M., Serrano J.C., Granado-Serrano A., Santamaría M., Yeramian N., Zorzano-Martínez M., Mora C, & Lecube A. (2023). PKN1 Kinase: A Key Player in Adipocyte Differentiation and Glucose Metabolism. Nutrients, 15(10), 2414.

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Neurodegeneration Assay Reagents

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Rotenone (R8875), sodium dodecyl sulfate (SDS, L3771), N, N-dimethylformamide (DMF, D4551), Dimethyl sulfoxide (DMSO, D8418), Phenylmethanesulfonyl fluoride (PMSF, 78830), Ethylenediaminetetraacetic acid (EDTA, E9884), Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA, E3889) and dopamine hydrochloride (H8502) were purchased from Sigma-Aldrich. Goat anti-mouse (115-035-003) and goat anti-rabbit (111-035-003) antibodies were purchased from Jackson ImmunoResearch Laboratories, INC. Alexa Fluor 488 conjugated goat anti-rabbit secondary antibody (A-11008) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, M-6494) were purchased from Thermo Fisher Scientific Inc. Anti-tyrosine hydroxylase antibody (anti-TH, AB152) was purchased from Merck Millipore. Anti-α-synuclein (610787) was purchased from BD Transduction Laboratories. Anti-β-actin (sc-47778) was purchased from Santa Cruz Biotechnology.

Liu L.F., Song J.X., Lu J.H., Huang Y.Y., Zeng Y., Chen L.L., Durairajan S.S., Han Q.B, & Li M. (2015). Tianma Gouteng Yin, a Traditional Chinese Medicine decoction, exerts neuroprotective effects in animal and cellular models of Parkinson’s disease. Scientific Reports, 5, 16862.

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Antibody-Based Signaling Pathway Analysis

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The antibody against LILRB4 was purchased from GeneTex (GTX33296). The antibodies against p-IkBα (9246), IkBα (4814), p65 (4764), p-p65 (3033), p-Ikkβ (2078), Ikkβ (8943), and GAPDH (2118) were purchased from Cell Signaling Technology. The goat anti-mouse (115-035-003) and goat anti-rabbit (111-035-003) secondary antibodies were purchased from Jackson Laboratory. The BCA protein assay kit was purchased from Pierce. Fetal calf serum (FCS) was obtained from HyClone. The cell culture reagents and all other reagents were obtained from Sigma.

Qiu T., Zhou J., Wang T., Chen Z., Ma X., Zhang L, & Zou J. (2019). Leukocyte immunoglobulin-like receptor B4 deficiency exacerbates acute lung injury via NF-κB signaling in bone marrow-derived macrophages. Bioscience Reports, 39(6), BSR20181888.

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Protein Extraction and Western Blot Analysis

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Proteins were extracted using RIPA lysis buffer containing a protease inhibitor cocktail (Sigma, St. Louis, MO, U.S.A.) and a phosphatase inhibitor cocktail (Roche, Mannheim, Germany). After quantitation using the Bradford method, protein lysates were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride membrane (Millipore). The membranes were blocked with nonfat milk for 1 h and incubated with primary antibodies, which are listed in Supplementary Table S1, overnight at 4°C. They were then incubated with goat anti-rabbit (111-035-003, Jackson) or anti-mouse (115-035-003, Jackson) HRP–conjugated secondary antibodies. Protein bands were visualized by enhanced chemiluminescence (Millipore). β-actin was used as a loading control.

Zhao Y., Yu Y., Zhao W., You S., Feng M., Xie C., Chi X., Zhang Y, & Wang X. (2019). As a downstream target of the AKT pathway, NPTX1 inhibits proliferation and promotes apoptosis in hepatocellular carcinoma. Bioscience Reports, 39(6), BSR20181662.

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