Secondary anti mouse and anti rabbit antibodies

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Secondary anti-mouse and anti-rabbit antibodies are laboratory reagents used to detect the presence of primary antibodies that have been raised against mouse or rabbit antigens. These secondary antibodies are conjugated with enzymes or fluorescent dyes, which allow for the visualization and quantification of the primary antibodies in various experimental techniques, such as Western blotting, immunohistochemistry, and ELISA.

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Lab products found in correlation

Pvdf membrane, by Bio-Rad (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) P erk, by Cell Signaling Technology (2 mentions) Transfection reagent, by Thermo Fisher Scientific (1 mentions) Puromycin, by Merck Group (1 mentions) Nicotinamide adenine dinucleotide phosphate, by Merck Group (1 mentions) Ab7817, by Abcam (1 mentions) Ab7778, by Abcam (1 mentions) Ecl plus reagent, by Bio-Rad (1 mentions) Oxidized glutathione, by Merck Group (1 mentions) Ripa regent, by Beyotime (1 mentions) β actin a5441, by Merck Group (1 mentions) Glutathione (gsh), by Merck Group (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Antibodies against c jun, by Cell Signaling Technology (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Cyt c, by Cell Signaling Technology (1 mentions) Anti bax, by Cell Signaling Technology (1 mentions) Anti bcl 2, by Cell Signaling Technology (1 mentions) Caspase 8, by Cell Signaling Technology (1 mentions)

27 protocols using secondary anti mouse and anti rabbit antibodies

Western Blot Analysis of Apoptotic Markers

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Five micrograms of total proteins were extracted with Pierce's M-Permammalian protein extraction reagent or 1 percent SDS and run on a 4 percent acrylamide concentration gel and a 10 percent acrylamide separation gel before being transferred to a Hybond P membrane (AP-Biotech). Membranes were pre-hybridized in PBS containing 0.1 percent (v/v) Tween 20 and 5% fat free milk (PBS-T). After that, the primary antibody was used to hybridize in PBS-T containing 5% fat free milk. Membranes were washed three times in PBS-T for five minutes each time and twice for ten minutes each time before being hybridized with secondary antibody in PBS-T containing 5% fat free milk.
Membranes were washed three times in PBS-T for five minutes each time and twice for ten minutes each time. ECL-plus Western blot detection reagent was used to visualize proteins of interest (AP-Biotech). Secondary antirabbit and anti-mouse antibodies (Santa Cruz) were used in conjunction with polyclonal rabbit anti-pro-caspase 9 and anti-pro-caspase 8 antibodies (Pharmingen), a polyclonal mouse antiGSTP1-1 antibody (Transduction Laboratories), a monoclonal mouse anti-b-actin (Sigma), a monoclonal mouse anti-PARP (Pharmingen), and secondary anti-rabbit and antimouse antibodies (Santa Cruz) were used.

Joicea A., Bashini J.M., Jose D., Kumar P., Munuswamy E., & Satheesh D. (2022). Curcumin (Curcuma longa): An Ecofriendly and Ayurvedic Antiproliferative Agent to Control Human Breast Cancer and Prostate Cancer. International Journal of Pharmaceutical Sciences Review and Research, 72(1).

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Integrin-Mediated Apoptosis Signaling

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Transfection reagents were purchased from Invitrogen (Carlsbad, CA). Puromycin was obtained from Sigma (St. Louis, MO). Antibodies were purchased as follows: anti-human β4 and α6 integrins from Abcam (Cambridge, MA); anti-mouse β4-integrin from R&D (Minneapolis, MN); anti-PARP, anti-Caspase-3, anti-Bcl-2 antibodies, and anti-mouse and anti-rabbit secondary antibodies from Santa Cruz Biotechnology (Santa Cruz, CA); anti-Bcl and -xL, anti-Bax and anti-Puma from Cell Signaling (Danvers, MA). Anti-β4-integrin (450-11A) antibody for immunohistochemistry was a kind gift from Dr. Rita Falcioni (Regina Elena Cancer Institute, Italy).

Zhang W., Zhang B., Vu T., Yuan G., Zhang B., Chen X., Manne U, & Datta P.K. (2017). Molecular characterization of pro-metastatic functions of β4-integrin in colorectal cancer. Oncotarget, 8(54), 92333-92345.

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Investigating EGCG's Therapeutic Potential

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EGCG (>99%) purified from green tea was obtained from Ebeikar Tea & Extracts Co., Ltd. (Hangzhou, China). Glutathione, oxidized Glutathione, GR (from baker yeast), and nicotinamide adenine dinucleotide phosphate (NADPH) were all obtained from Sigma (St. Louis, MO, USA). RIPA regent and BCA protein assay kit were products of Beyotime Biotechnology (Shanghai, China). Anti-mouse and anti-rabbit secondary antibodies were obtained from Santa Cruz (Dallas, TX, USA). The primary antibodies against UT-A1 (SAB5200108) and β-actin (A5441) were purchased from Sigma (St. Louis, MO, USA). The primary antibodies against AQP2 (#3487), Bax (#2772), p53 (#9282) and caspase3 (#9662) were purchased from Cell Signal Technology, Inc. (Boston, MA, USA). The primary antibodies against CTGF (ab6992), α-SMA (ab7817) and collagen III (ab7778) were products of Abcam (Cambridge, UK). The primary antibody against PCNA (BM0104) was purchased from Boster Biological Technology Co., Ltd. (Wuhan, China). ECL Plus reagent and PVDF membrane were purchased from Bio-Rad Laboratories, Inc. (Hercules, CA, USA). Other chemicals were of the highest grade available.

Wang X., Yang L., Wang J., Zhang Y., Dong R., Wu X., Yang C.S., Zhang Z, & Zhang J. (2019). A mouse model of subacute liver failure with ascites induced by step-wise increased doses of (-)-epigallocatechin-3-gallate. Scientific Reports, 9, 18102.

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Protein Signaling Pathway Analysis

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IL-6, soluble IL-6 receptor, and CYR61 protein were purchased from PeproTech, Inc. (Rocky Hill, NJ, USA). Monoclonal antibodies against human extracellular signal-regulated kinases 1/2 (ERK 1/2), phospho-ERK 1/2, Cyr61, beta-actin, and early growth response protein 3 (EGR3), Nuclear receptor subfamily 4 group A member 1 (NR4A1), Activating Transcription factor 3 (ATF3) as well as anti-mouse and anti-rabbit secondary antibodies were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Antibodies against c-Jun were purchased from Cell Signaling (Danvers, MA, USA). Cyr61-neutralising antibody was purchased from Novus Biologicals (Littleton, CO, USA).

Choi C., Jeong W., Ghang B., Park Y., Hyun C., Cho M, & Kim J. (2020). Cyr61 synthesis is induced by interleukin-6 and promotes migration and invasion of fibroblast-like synoviocytes in rheumatoid arthritis. Arthritis Research & Therapy, 22, 275.

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Apoptosis Pathway Assays in Cells

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Rotenone, vanillin, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2-7-diacetyl dichlorofluorescein (DCFH-DA), rhodamine 123 (Rh-123), heat-inactivated fetal bovine serum (FBS), Dulbecco's modified Eagle's medium (DMEM), antibiotic/antimycotic, EDTA, and Trypsin-EDTA were procured from Sigma Chemicals Co. (St. Louis, USA). Anti-Bcl-2, anti-Bax, caspase-3, caspase-8, caspase-9, cyt-c, and anti-JNK and anti-P38 MAPK antibodies were obtained from Cell Signaling (USA) and β-actin, anti-mouse, and anti-rabbit secondary antibodies were purchased from Santa Cruz Biotechnology, Inc. (USA).

Dhanalakshmi C., Manivasagam T., Nataraj J., Justin Thenmozhi A, & Essa M.M. (2015). Neurosupportive Role of Vanillin, a Natural Phenolic Compound, on Rotenone Induced Neurotoxicity in SH-SY5Y Neuroblastoma Cells. Evidence-based Complementary and Alternative Medicine : eCAM, 2015, 626028.

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Epithelial-Mesenchymal Transition Modulation

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The HCC cell lines Huh7 and HepG2 were purchased from the American type culture collection (Manassas, VA, USA) and cultured in DMEM containing 10% fetal bovine serum at 37 °C in a 5% CO₂ atmosphere. Polyclonal antibodies against N-cadherin, vimentin, slug, twist, snail, and β-actin were purchased from Genetex (Irvine, CA, USA) and Cell Signaling Technology (Beverly, MA, USA). The anti-mouse and anti-rabbit secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The corylin powder (purity above 98% as measured by HPLC) was purchased from Shanghai BS Bio-Tech Co., Ltd. (Shanghai, China) and dissolved in DMSO to a final concentration of 100 mM. Commercial si-GAS5 and negative-control siRNA were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Chen C.Y., Chen C.C., Shieh T.M., Hsueh C., Wang S.H., Leu Y.L., Lian J.H, & Wang T.H. (2018). Corylin Suppresses Hepatocellular Carcinoma Progression via the Inhibition of Epithelial-Mesenchymal Transition, Mediated by Long Noncoding RNA GAS5. International Journal of Molecular Sciences, 19(2), 380.

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Western Blot Analysis of Cell Lysates

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Whole cells were lysed in RIPA buffer (150 mM NaCl, 1% NP-40, 0.5% Na deoxycholate, 0.1% SDS, 50 mM Tris-Cl, pH = 8.0) supplemented with a protease inhibitor cocktail consisting of 1 μg/mL aprotinin, 1 μg/mL leupeptin, 3 μg/mL Pepstatin, 1 mM NaVO3, 1 mM NaF, 0.5 μM DTT (Sigma Chemical, St. Louis, MO). Protein lysates were separated on 10% SDS-polyacrylamide gels. Antibodies used were as follows: cFLIP human (Enzo Life Sciences #ALX-804-961-0100, Farmingdale, NY) [16 (link)]; cFLIP mouse (Abcam #ab8421, Cambridge, MA); total PARP, cleaved PARP, caspase 8, TRAIL-R2, p53, and NFKB1 (Cell Signaling Technology #9532, #5625, #9746, #8074, #2524, and #13586 Danvers, MA); TRAIL-R1 (Thermo Scientific, #32A1380); E-cadherin (BD Transduction Laboratories #610182, Franklin Lakes, NJ); β-actin (Sigma Chemical #A5441, St. Louis, MO); GAPDH (Life Technologies #AM4300, Carlsbad, CA); anti-mouse and anti-rabbit secondary antibodies (Santa Cruz Biotechnology #sc2005 and #sc-2004, Dallas, TX). Chemiluminescent western blot analyses were performed using ECL detection (EMD Millipore, Darmstadt, Germany).

Padmanabhan C., Rellinger E.J., Zhu J., An H., Woodbury L.G., Chung D.H., Waterson A.G., Lindsley C.W., Means A.L, & Beauchamp R.D. (2017). cFLIP critically modulates apoptotic resistance in epithelial-to-mesenchymal transition. Oncotarget, 8(60), 101072-101086.

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Protein Expression Analysis in Cells

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After the incubation period, cells were washed with ice-cold phosphate buffered saline (PBS) and immediately frozen by placing the dishes in fluid nitrogen. Lysates were prepared as described previously using lysis buffer P and subjected to SDS-PAGE [19 (link)]. Membranes were probed with antibodies directed against actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), EGFR (Santa Cruz Biotechnology), HIF-1α (BD Transduction Laboratories, San Jose, CA, USA), BNIP3 (Abcam, Cambridge, UK), and phosphorylated (P-) p90RSK, P-Akt, P-p42/44 MAP kinase (MAPK), P-ribosomal protein S6 (P-RPS6) as well as the eukaryotic translation initiation factor 4E (eIF4E9) as a loading control using the PathScan Multiplex Western Cocktail I (Cell Signaling Technology, Danvers, MA, USA, #7100). The secondary anti-rabbit and anti-mouse antibodies were purchased from Santa Cruz Biotechnology. Chemiluminescence solution was used for detection [25 (link)].

Urban H., Maurer G.D., Luger A.L., Lorenz N.I., Sauer B., Stroh C., Trojan J., Mittelbronn M., Steinbach J.P., Harter P.N, & Ronellenfitsch M.W. (2020). Cetuximab-Mediated Protection from Hypoxia- Induced Cell Death: Implications for Therapy Sequence in Colorectal Cancer. Cancers, 12(10), 3050.

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EECM Effects on Osteoblast Differentiation

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EECM was provided by the Korea Institute Science Technology Natural Products Library (Gangneung, South Korea). Alpha minimum essential medium (α-MEM), Dulbecco’s modified Eagle’s medium, penicillin-streptomycin, and phosphate buffered saline (PBS) were purchased from Welgene (Gyeongsan, South Korea). Fetal bovine serum was purchased from Gibco, Gaithersburg, MD. Antibodies against β-actin, Runx2, OPG, OSX, BMP2/4, ALP, and secondary anti-rabbit and anti-mouse antibodies were purchased from Santa Cruz Biotechnology, Dallas, TX. Antibodies against phospho-extracellular signal-regulated protein kinase (p-ERK), ERK, and COL1A1 were purchased from Cell Signaling Technology (Danvers, MA). Alizarin Red S (ARS), 1,25-dihydroxyvitamin D3, L-ascorbic acid, β-glycerophosphate, ICI 182,780 and genistein were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO). Ultra Pure bovine serum albumin (BSA) was purchased from GenDEPOT (Katy, TX). Noggins was purchased from Peprotech (London, U.K.).

Park J.H., Son Y.J., Lee C.H., Nho C.W, & Yoo G. (2020). Circaea mollis Siebold & Zucc. Alleviates postmenopausal osteoporosis in a mouse model via the BMP-2/4/Runx2 pathway. BMC Complementary Medicine and Therapies, 20, 123.

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Western Blot Analysis of Claudin-1 Knockdown and TGF-β Signaling

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Proteinase and phosphatase inhibitors cocktail were from Sigma-Aldrich (St. Louis, MO, USA). PVDF membrane (1620177), Precision Plus Protein™ Dual Color Standards (1610394) and ECL reagents (1705061) were from Bio-Rad Laboratories (Hercules, CA, USA). Cells treated by siRNA knockdown of claudin 1 or treated with TGF-β (with/without inhibitors) and western blotting was conducted as previously published [20 (link)]. GAPDH was probed as loading control for each individual experiment, representative results from at least two experimental replicates are shown. Primary antibodies are listed in Table 1; secondary anti-rabbit and anti-mouse antibodies were from Santa Cruz Biotechnology (Dallas, TX, USA). Dilutions for primary and secondary antibodies were 1:1000 and 1:5000 respectively. Quantification of band intensity was measured using Adobe PhotoShop v. 21.1.1 (Adobe Systems, San Jose, CA, USA).

Wang K., Pascal L.E., Li F., Chen W., Dhir R., Balasubramani G.K., DeFranco D.B., Yoshimura N., He D, & Wang Z. (2020). Tight junction protein claudin 1 is down-regulated by TGF-β1 via MEK signaling in benign prostatic epithelial cells. The Prostate, 80(14), 1203-1215.

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