Secondary goat anti rabbit and goat anti mouse antibodies

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Secondary goat anti-rabbit and goat anti-mouse antibodies are used as detection reagents in various immunoassays and immunohistochemical techniques. They specifically recognize and bind to primary antibodies raised in rabbit or mouse, enabling the visualization and amplification of target analytes or cellular structures.

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Secondary goat anti-mouse and anti-rabbit antibodies HRP-conjugated goat anti-mouse and anti-rabbit secondary antibodies Horseradish peroxidase (HRP)-conjugated rabbit anti-mouse and goat anti-rabbit secondary antibodies Horseradish peroxidase-conjugated goat anti-rabbit and goat anti-mouse secondary antibodies Goat anti-rabbit and goat anti-mouse IgG horseradish peroxidase-conjugated secondary antibodies Secondary anti-rabbit and anti-goat antibodies Secondary anti-mouse and anti-rabbit antibodies Anti-goat secondary antibodies Anti-mouse secondary antibodies Goat anti-rabbit antibodies

7 protocols using secondary goat anti rabbit and goat anti mouse antibodies

Western Blot Analysis of Protein Expressions

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Tissues or cells were solubilized in cold RIPA lysis buffer. Proteins were separated with 12% SDS-PAGE, and transferred onto a polyvinylidene difluoride (PVDF) membrane. The membrane was incubated with TBST containing 5% skimmed milk at 37°C for 2 h. The membrane was then incubated with rabbit anti-ROS, rabbit anti-FIG, and mouse anti-GAPDH primary antibodies (all from Santa Cruz Biotechnology, Inc., Santa Cruz, USA), respectively, at room temperature for 2 h. After washing with PBST 4 times for 10 min each time, the membrane was incubated with the goat anti-rabbit and goat anti-mouse secondary antibodies (Santa Cruz Biotechnology, Inc.) at 4°C overnight. After washing with PBST 4 times for 10 min each time, an ECL kit (Pierce Chemical, Rockford, IL, USA) was used to perform chemiluminent detection. Image-Pro plus software 6.0 was used to analyze the relative protein expression, represented as the density ratio versus GAPDH. GAPDH was used as an internal reference.

DENG G., HU C., ZHU L., HUANG F., HUANG W., XU H, & NIE W. (2014). Downregulation of ROS-FIG inhibits cell proliferation, colony-formation, cell cycle progression, migration and invasion, while inducing apoptosis in intrahepatic cholangiocarcinoma cells. International Journal of Molecular Medicine, 34(3), 661-668.

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Western Blot Analysis of PARP-1 Cleavage

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For Western blotting, cells were homogenized in RIPA buffer (Cell Signaling, Danvers, MA) plus phosphatase and protease inhibitor cocktail, 10 mM NaF, 1 mM PMSF and 1 mM Na3VO4. Protein extracts were purified, and concentrations were measured with Pierce Protein Assay kit (Thermo Fisher Scientific). The following antibodies were used in this study: Anti-MTAP antibody (Rabbit polyclonal, 1:2000) from ProteinTech, anti-poly (ADP-ribose) polymerase-1 (PARP-1) antibody (Rabbit monoclonal, 1:2000) from Cell Signaling (Danvers, MA), and β-actin (Mouse monoclonal, 1:10000) from MilliporeSigma (Burlington, MA). We assessed the cleavage of PARP-1 using Western blot. PARP-1 cleavage by caspase-3 and −7 during apoptosis into a 24 kDa N-terminal fragment and an 89 kDa fragment has been used as a marker of apoptotic animal cells^{54 (link)}. Goat anti-rabbit and goat anti-mouse secondary antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX). The chemiluminescence signals were developed with Bio-Rad ChemiDoc XRS + System (Bio-Rad, Hercules, CA).

Alhalabi O., Chen J., Zhang Y., Lu Y., Wang Q., Ramachandran S., Tidwell R.S., Han G., Yan X., Meng J., Wang R., Hoang A.G., Wang W.L., Song J., Lopez L., Andreev-Drakhlin A., Siefker-Radtke A., Zhang X., Benedict W.F., Shah A.Y., Wang J., Msaouel P., Zhang M., Guo C.C., Czerniak B., Behrens C., Soto L., Papadimitrakopoulou V., Lewis J., Rinsurongkawong W., Rinsurongkawong V., Lee J., Roth J., Swisher S., Wistuba I., Heymach J., Wang J., Campbell M.T., Efstathiou E., Titus M., Logothetis C.J., Ho T.H., Zhang J., Wang L, & Gao J. (2022). MTAP deficiency creates an exploitable target for antifolate therapy in 9p21-loss cancers. Nature Communications, 13, 1797.

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Integrin Signaling Pathway Antibodies

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Anti-FKBP10 (PA5-63387, 0.4 µg/mL) was purchased from Invitrogen (Rockford, IL, USA). Anti-AKT (9272,1:1000), anti-p-AKT₃₀₈(4056,1:1000), anti-p-AKT₄₇₃(9271L,1:1000), anti-integrin αV (4711,1:500) and α5 (4705,1:1000), anti-GAPDH (2118,1:1000) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-integrin α6 (ab97760,1:500) was purchased from Abcam (Cambridge, MA, USA). Anti-integrin α1 (mab5676, 2µg/mL) and α2 (mab12331,1µg/mL) were purchased from the R&D system (Minneapolis, MN, USA).
Secondary goat anti-rabbit and goat anti-mouse antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Matrigel was purchased from Corning (Corning Life Science, Tewksbury, MA, USA).

Gong L.B., Zhang C., Yu R.X., Li C., Fan Y.B., Liu Y.P, & Qu X.J. (2020). FKBP10 Acts as a New Biomarker for Prognosis and Lymph Node Metastasis of Gastric Cancer by Bioinformatics Analysis and in Vitro Experiments. OncoTargets and therapy, 13, 7399-7409.

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Gastric Cancer Cell Lines Characterization

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The gastric cancer cell lines BGC823, MGC803, HGC27 and AGS were purchased from the Chinese Academy of Sciences (Shanghai, China). MKN7 was from the Japanese Collection of Research Bioresources (JCRB Cell Bank, Osaka, Japan). SNU216 was obtained from the Korea Cell Line Bank (KCLB, Seoul, Korea). All cell lines except for AGS maintained in F-12K medium, were cultured in RPMI-1640 medium added with 10% heat-inactivated FBS. They were incubated at 37°C in saturated humidity with 5% CO2. Rabbit anti-FTO (#31687), anti-integrin β1 (#9699S), anti-GAPDH (#2118) and anti-LAMC1 (#4577) were obtained from Cell Signaling Technology (Danvers, MA, USA). Secondary goat anti-rabbit and goat anti-mouse antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Cycloleucine was purchased from Sigma (Sigma-Aldrich Co., St Louis, MO, USA). Matrigel was obtained from Corning (Corning Life Science, Tewksbury, MA, USA).

Wang D., Qu X., Lu W., Wang Y., Jin Y., Hou K., Yang B., Li C., Qi J., Xiao J., Che X, & Liu Y. (2021). N6-Methyladenosine RNA Demethylase FTO Promotes Gastric Cancer Metastasis by Down-Regulating the m6A Methylation of ITGB1. Frontiers in Oncology, 11, 681280.

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Adriamycin Sensitivity in SGC7901 Cells

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The human GC cell line SGC7901 was obtained from the Academy of Military Medical Science (Beijing, China). The Adr -resistant variant of SGC7901 (SGC7901/Adr) was kindly provided by the Fourth Military Medical University (Xi'an, China). Antibodies for SPARC were obtained from Cell Signaling Technology (Beverly, MA, USA), and the secondary goat anti-rabbit and goat anti-mouse antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Adriamycin sensitivity of SGC7901 with or without SPARC siRNA transfection was further investigated using MTT assay. The detailed information and the techniques for cell culture, Western Blot, siRNA, and MTT assay were described in our previous study elsewhere [20 (link), 47 (link)] and the current Supplementary File S1.

Li Z., Li A.D., Xu L., Bai D.W., Hou K.Z., Zheng H.C., Qu X.J, & Liu Y.P. (2016). SPARC expression in gastric cancer predicts poor prognosis: Results from a clinical cohort, pooled analysis and GSEA assay. Oncotarget, 7(43), 70211-70222.

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Comprehensive Protein Expression Analysis

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Western blot was performed as previously described.14 (link) The following antibodies were used: Anti-RANK antibody was obtained from Bethyl Laboratories, Inc. (Montgomery, TX, USA). Anti-actin antibody was produced by Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Anti-EGFR, anti-phospho-EGFR (Tyr1068), anti-AKT, anti-p-AKT (Ser473), anti-ERK1/2, anti-p-ERK1/2 (Thr202/Tyr204), anti-c-Src, and anti-p-c-Src (Y416) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Secondary goat anti-rabbit and goat anti-mouse antibodies were purchased from Santa Cruz Biotechnology Inc.

Zhang X., Song Y., Song N., Zhang Y., Zhang L., Wang Y., Wang Z., Qu X, & Liu Y. (2017). RANKL/RANK pathway abrogates cetuximab sensitivity in gastric cancer cells via activation of EGFR and c-Src. OncoTargets and therapy, 10, 73-83.

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Western Blot Analysis of Protein Expression

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Western blot analysis was conducted as previously described [32 (link)], in samples collected 48 hours after transfection. The following primary antibodies were used: CD36 (R&D, Minneapolis, USA), DEK (Abcam), c-myc (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), phospho-ERK, ERK, E-cadherin, Vimentin, Snail, ZEB1, GSK-3β, p-Ser9-GSK-3β, β-catenin, phospho-β-catenin, and GAPDH (Cell Signaling Technology). Secondary goat anti-rabbit and goat anti-mouse antibodies were purchased from Santa Cruz Biotechnology. The corresponding catalog numbers and dilutions are shown in Supplementary Table 1.

Wang J., Wen T., Li Z., Che X., Gong L., Jiao Z., Qu X, & Liu Y. (2020). CD36 upregulates DEK transcription and promotes cell migration and invasion via GSK-3β/β-catenin-mediated epithelial-to-mesenchymal transition in gastric cancer. Aging (Albany NY), 13(2), 1883-1897.

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