Mouse anti mmp9

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Mouse anti-MMP9 is a monoclonal antibody that specifically binds to and detects Matrix Metalloproteinase 9 (MMP9), an enzyme involved in the degradation of extracellular matrix proteins.

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Lab products found in correlation

8 protocols using mouse anti mmp9

Lentiviral shRNA Knockdown Constructs

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Scrambled, TLK1-, and MK5-specific shRNA constructs in lentiviral GFP vector were purchased from Origene (Rockville, MD, USA, TLK1 shRNA cat# TL320623, MK5 shRNA cat# TL320583, scrambled shRNA cat# TR30021). The following primary antibodies were used in this study: rabbit anti-TLK1 (ThermoFisher, Waltham, MA, USA, cat# 720397), mouse anti-PRAK/MK5 (Santa Cruz Biotechnology, Dallas, TX, USA, cat# sc-46667), rabbit anti Phospho- FAK Y397 (ThermoFisher, cat# 44-624G), rabbit anti-Phospho- FAK Y861 (abcam, Cambridge, MA, USA, cat# ab4804), rabbit anti-FAK (Cell signaling technology, Danvers, MA, USA, cat# 3285S), rabbit anti-phospho-paxillin Y118 (Cell signaling technology, cat# 2541S), rabbit anti-paxillin (Santa Cruz Biotechnology, cat# sc-5574), rabbit anti-phospho-HSP27 S82 (ThermoFisher, cat# 44-534G), mouse anti-HSP27 (ThermoFisher, cat# MA3-015), rabbit anti- ERK3 (Cell signaling technology, cat# 4067S), mouse anti-MMP2 (Santa Cruz Biotechnology, cat# sc-13595), mouse anti-MMP9 (Santa Cruz Biotechnology, cat# sc-13520), mouse anti-MMP3/10 (Santa Cruz Biotechnology, cat# sc-374029), mouse anti-Ki-67 (Cell signaling technology, cat# 9449S), and rabbit anti-GAPDH (Cell signaling technology, cat# 2118S).

Khalil M.I, & De Benedetti A. (2022). The TLK1–MK5 Axis Regulates Motility, Invasion, and Metastasis of Prostate Cancer Cells. Cancers, 14(23), 5728.

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Antibody-Based Analysis of Signaling Pathways

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The antibodies used in this study included the following: anti-IL-6 neutralizing antibody (R&D systems), rabbit anti-JAK2, rabbit anti-phospho (Tyr1007-10008)-JAK2, rabbit anti-STAT3, rabbit anti-phospho (Tyr705)-STAT3, rabbit anti-Vimentin, rabbit anti-Snail (Cell Signaling); mouse anti-Twist, mouse anti-MMP9, goat anti-β-actin, mouse anti-HSC70 (Santa Cruz); peroxidase affinipure anti-mouse IgG and peroxidase affinipure anti-rabbit IgG (Jackson ImmunoResearch).

Bouaouiche S., Ghione S., Sghaier R., Burgy O., Racoeur C., Derangère V., Bettaieb A, & Plenchette S. (2021). Nitric Oxide-Releasing Drug Glyceryl Trinitrate Targets JAK2/STAT3 Signaling, Migration and Invasion of Triple-Negative Breast Cancer Cells. International Journal of Molecular Sciences, 22(16), 8449.

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Molecular Markers in Neurodegeneration

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We used the following primary antibodies: rabbit anti-mBdnf and proBdnf (Santa Cruz Biotechnology, sc-546), rabbit anti phospho-MEK1/2 (Ser217/221) (Cell Signaling Technology, #9121), rabbit anti MEK1/2 (Cell Signaling Technology, #9122), rabbit anti-Actin (A2066; Sigma-Aldrich), rabbit anti-Hprt (ab10479; Abcam), rabbit anti-Gapdh (Cell Signaling; 2118), HRP anti-rabbit (P0448; Dako), mouse anti-Mmp7 (Santa Cruz; sc-515703), mouse anti-Mmp9 (Santa Cruz; sc-393859), mouse anti-Mmp3 (Santa Cruz; sc-21732), mouse anti-Furin (Santa Cruz; sc-133142), mouse anti-Plasminogen (Santa Cruz; sc-376324), mouse anti-Timp1 (Santa Cruz; sc-365905), rabbit anti-PC1/3 (Cell Signaling; 11914), IRDye 800CW anti-mouse (LI-COR; 925–32210) and IRDye 680RD anti-rabbit (LI-COR; 925–68071)

Spychala A, & Rüther U. (2019). FTO affects hippocampal function by regulation of BDNF processing. PLoS ONE, 14(2), e0211937.

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Epithelial-Mesenchymal Transition Protein Analysis

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Cultured cells were collected and solubilized using protein lysis buffer. The proteins were then separated by size using SDS-PAGE and transferred to polyvinyl difluoride membranes (Millipore). The membranes were incubated with primary antibodies followed by secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA). The immunoreactive proteins were detected using an enhanced chemiluminescence kit (Millipore). The primary antibodies were rabbit anti- E-cadherin, rabbit anti-N-cadherin, rabbit anti-Vimentin (Cell Signaling Technology, Danvers, MA, USA), mouse anti-MMP9, mouse anti-MMP2, mouse anti-JNK1, mouse anti-p-JNK, rabbit anti-c-Jun, mouse anti-p-c-Jun, mouse anti-FAK, mouse anti-β-actin (Santa Cruz Biotechnology), rabbit anti-Paxillin, and rabbit anti-p-Paxillin (Abcam, Cambridge, MA, USA). The digital images of the Western blotting bands were quantified by Meta Morph software (MDS Analytical Technologies) after the background subtraction.

Cai J., Du S., Wang H., Xin B., Wang J., Shen W., Wei W., Guo Z, & Shen X. (2017). Tenascin-C induces migration and invasion through JNK/c-Jun signalling in pancreatic cancer. Oncotarget, 8(43), 74406-74422.

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Doxorubicin-Loaded Nanoparticle Delivery

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Doxorubicin (DOX), indocyanine green (ICG), bovine serum albumin (BSA), tetraethylorthosilicate (TEOS), and cetyltrimethylammonium bromide (CTAB) were obtained from Sigma-Aldrich (United States). 3,3′-Dioctadecyloxacarbocyanine perchlorate (Dio) and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) were bought from Biyuntian (China). Polyvinylidene fluoride membranes (PVDF) were acquired from Millipore (United States). MTT and Ki67 reagents were provided by Yeasen Corporation (China). RPMI 1640 medium, FBS, penicillin, and streptomycin were provided by Hyclone (United States). You Ning Wei Corporation (China) offered the other reagents, including acrylamide, mouse anti-MMP2 (Santa Cruz, United States), mouse anti-MMP9 (Santa Cruz, United States), rabbit anti-calnexin (Santa Cruz, United States), rabbit anti-CD81 (ProteinTech, Chicago, IL, United States), and rabbit anti-CD63 (Abcam, Cambridge, Britain).

Tian R., Wang Z., Niu R., Wang H., Guan W, & Chang J. (2020). Tumor Exosome Mimicking Nanoparticles for Tumor Combinatorial Chemo-Photothermal Therapy. Frontiers in Bioengineering and Biotechnology, 8, 1010.

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Western Blot Analysis of Hepatocellular Carcinoma

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Proteins were extracted from HCC tissues and cells using a cell lysis buffer. A total of 30 μg of protein was separated by 10% SDS-PAGE electrophoresis and transferred onto PVDF membranes (Amersham, Little Chalfont, UK). After blocking with Tween–Tris-buffered saline (T-TBS) containing 5% nonfat milk powder, the membranes were incubated with primary antibodies at 4°C overnight. The primary antibodies were utilized for Western blot: mouse anti-Rap2B, mouse anti-p-FAK, mouse anti-MMP-2, mouse anti-MMP-9, and mouse anti-GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA). Subsequently, the membranes were washed with TBST three times and incubated with peroxidase-conjugated goat anti-mouse IgG (1:5,000; Santa Cruz Biotechnology) for 3 h at room temperature. The target protein was visualized by enhanced chemiluminescence (Pierce, Rockford, IL, USA).

Zhang L., Duan H.B, & Yang Y.S. (2017). Knockdown of Rap2B Inhibits the Proliferation and Invasion in Hepatocellular Carcinoma Cells. Oncology Research, 25(1), 19-27.

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Western Blot Analysis of Angiogenic Factors

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Treated cells were lysed in ice-cold RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) containing ethylenediamine tetraacetic acid-free protease inhibitor cocktail (Roche Diagnostics). Equal amounts of samples (30 µg of protein) were separated on a NuPAGE 4-12% bis-tris gel (Invitrogen) and then transferred onto a nitrocellulose membrane. After blocking, membranes were incubated with primary antibodies, including rabbit anti-MMP2 (1:1,000, Santa Cruz Biotechnologies, Santa Cruz, CA, USA), mouse anti-MMP9 (1:1,000, Santa Cruz Biotechnologies), rabbit anti-TIMP1 (1:2,000, Abcam, Cambridge, MA, USA), mouse anti-TIMP2 (1:2,000, Abcam), rabbit anti-VEGF (1:1,000, Santa Cruz Biotechnologies), rabbit anti-CD31 (1:1,000, Abcam), and mouse anti-β-actin (1:1000, Abcam). After washing, the membrane was incubated with secondary antibody, and bands were visualized using ECL methods (Amersham, Arlington Height, IL, USA). Protein bands were detected using LAS-1000 (Fujifilm Medical Systems USA, Stamford, CT, USA).

Park Y.H., Jung A.R., Kim G.E., Kim M.Y., Sung J.W., Shin D., Cho H.J., Ha U.S., Hong S.H., Kim S.W, & Lee J.Y. (2019). GV1001 inhibits cell viability and induces apoptosis in castration-resistant prostate cancer cells through the AKT/NF-κB/VEGF pathway. Journal of Cancer, 10(25), 6269-6277.

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Resveratrol Modulates Bladder Cancer Cell Signaling

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Reagents. Resveratrol (purity, >99%) was purchased from Sigma-Aldrich; Merck Millipore (Darmstadt, Germany). The primary antibodies mouse anti-MMP-2 (catalog no. sc-13595), mouse anti-MMP-9 (catalog no. sc-21733), mouse anti-phosphorylated (p)-ERK1/2 (catalog no. sc-81492), mouse anti-ERK1/2 (catalog no. sc-135900), mouse anti-p-JNK1/2 (catalog no. sc-293137), mouse anti-JNK1/2 (catalog no. sc-7345) and mouse anti-β-actin (catalog no. sc-47778), and a horseradish peroxidase-conjugated goat anti-mouse secondary antibody (catalog no. sc-2031), were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). The bicinchoninic acid protein assay kit was purchased from Pierce; Thermo Fisher Scientific, Inc. (Waltham, MA, USA).
Cell culture. The T24 human bladder cancer cell line was obtained from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences (Shanghai, China). The cells were cultured in RPMI-1640 medium (HyClone; GE Healthcare, Logan, UT, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS; SAFC Biosciences Inc., Lenexa, KS, USA), 100 U⁄ml penicillin and 100 mg/l streptomycin. Cultures were maintained in a humidified atmosphere containing 5% CO 2 at 37˚C.

Bai Y., Yang H., Zhang G., Hu L., Lei Y., Qin Y., Yang Y., Wang Q., Li R, & Mao Q. (2017). Inhibitory effects of resveratrol on the adhesion, migration and invasion of human bladder cancer cells. Molecular medicine reports, 15(2).

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