Anti gapdh gtx627408

Manufactured by GeneTex

Sourced in United States

Anti-GAPDH (GTX627408) is a laboratory reagent used for the detection of the GAPDH protein. GAPDH is a key enzyme involved in the glycolysis pathway. This antibody can be used in various analytical techniques to identify and quantify the GAPDH protein in biological samples.

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GAPDH (204 citations) Anti-GAPDH (94 citations) GTX627408 (22 citations) GAPDH (GTX627408) (4 citations)

4 protocols using anti gapdh gtx627408

Immunoblot Analysis of Apoptosis Regulators

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CRC cells (4 × 10⁵) were treated with Obatoclax (0, 100, 200 μM) for 24 h, or treated with Obatoclax for 22 h and then combined with MG132 (20 μM) for an additional 2 h. After drug treatment, cells were lysed, subjected to SDS-PAGE, blot transfer, incubation with primary and secondary antibodies, exposure to enhanced chemiluminescence (ECL) reagent, and final visualization of the antigen-of-interest using the LAS3000 system (Fujifilm; Tokyo, JPN) as stated previously [45 (link),46 (link)]. Densitometry analysis of immunoblot signals was accomplished by ImageJ software (National Institute of Health, Bethesda, MD, USA). Anti-survivin (#ab469) and anti-TCF4 (#ab76151) antibodies were bought from Abcam (Cambridge, GBR). Anti-β-catenin (sc-7963) and anti-c-MYC (sc-40) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-LEF1 (GTX129186) and anti-GAPDH (GTX627408) were obtained from GeneTex (Hsinchu, TWN), and anti-β-tubulin (#T2200) was bought from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies against cyclin D1 (#2926), HA-tag (#3724), and cleaved forms of caspases-3 (#9664), -8 (#9496), -9 (#9501), and PARP (#9541) were all purchased from Cell signaling Technology (Boston, MA, USA).

Or C.H., Huang C.W., Chang C.C., Lai Y.C., Chen Y.J, & Chang C.C. (2020). Obatoclax, a Pan-BCL-2 Inhibitor, Downregulates Survivin to Induce Apoptosis in Human Colorectal Carcinoma Cells Via Suppressing WNT/β-catenin Signaling. International Journal of Molecular Sciences, 21(5), 1773.

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Extracellular Vesicle Protein Detection

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TGFβ1 was purchased from Peprotech (Rocky Hill, NJ). The primary antibodies were used at 1:1000 ratio. Monoclonal anti-CD63 (H5C6, The Developmental Studies Hybridoma Bank, Iowa City, IA), anti-CD81 (MAB 4615, R&D Systems, Minneapolis, MN), anti-Thy-1.2 (HO-13-4, ATCC and 550402, BD Biosciences, San Diego, CA), anti-GAPDH (GTX627408, GeneTex, Irvine, CA) and rabbit polyclonal anti-calnexin (2679S, Cell Signaling, Danvers, MA) were used to detect EV and cellular proteins. Monoclonal anti-FN-EDA (ab6328, Abcam, Cambridge, MA), monoclonal anti-collagen I (GTX26308, GeneTex) and rabbit polyclonal anti-α-SMA (CBL171-I, EMD Millipore, Temecula, CA) anti-collagen III (GTX111643, GeneTex) and anti-N-cadherin (GTX127345, GeneTex) were used in western blotting studies.

Shentu T.P., Huang T.S., Cernelc-Kohan M., Chan J., Wong S.S., Espinoza C.R., Tan C., Gramaglia I., van der Heyde H., Chien S, & Hagood J.S. (2017). Thy-1 dependent uptake of mesenchymal stem cell-derived extracellular vesicles blocks myofibroblastic differentiation. Scientific Reports, 7, 18052.

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Quantification of Viral Titers and Proteins

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Lung tissues were homogenized in DMEM using the bead-beating technology (Precellys Lysing kits; Bertin Technologies, Aix-en-Provence, France). Following centrifugation, supernatants were stored in small aliquots. Viral titers were determined using a plaque assay following a previously described methodology [12 (link)]. For western analysis, thawed supernatants were mixed with equal amounts of RIPA buffer containing lysozyme and proteinase inhibitors. The protein content was measured with a protein assay kit (Bio-Rad, Richmond, CA, USA). Equal amounts of cellular proteins were separated by SDS-PAGE and subsequently transferred onto polyvinylidene fluoride membranes. Subsequently, the membranes were immunoblotted with specific primary antibodies and subsequently exposed to horseradish peroxidase (HRP)-conjugated secondary antibodies. The Immobilon Western HRP Chemiluminescence Substrate (Millipore, Burlington, MA, USA) was used for blot development. Mouse monoclonal anti-nucleocapsid (GTX632269, 1:2000 dilution) and anti-GAPDH (GTX627408, 1:5000 dilution) antibodies were obtained from Genetex (Hsinchu, Taiwan).

Chin Y.F., Tang W.F., Chang Y.H., Chang T.Y., Lin W.C., Lin C.Y., Yang C.M., Wu H.L., Liu P.C., Sun J.R., Hsu S.C., Lee C.Y., Lu H.Y., Chang J.Y., Jheng J.R., Chen C.C., Kau J.H., Huang C.H., Chiu C.H., Hung Y.J., Tsai H.P, & Horng J.T. (2022). Orally delivered perilla (Perilla frutescens) leaf extract effectively inhibits SARS-CoV-2 infection in a Syrian hamster model. Journal of Food and Drug Analysis, 30(2), 252-270.

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Western Blotting of Protein Targets

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Proteins levels were analyzed by Western blotting using a standard protocol^{59 (link)}. Antibodies used in this study were: anti-Rab7 (9367S, 1:1000), anti-Alix (2171S, 1:1000), anti-CD81 (10037S, 56039S, 1:1000), anti-Tom20 (42406S, 1:1000), anti-Rab4 (2167S, 1:1000), anti-Rab5 (3547S, 1:1000), anti-Rab9 (5118S, 1:1000), anti-Rab11 (5589S, 1:1000), anti-LC3A/B (4108S, 1:1000), anti-Rab27 (69295S, 1:1000), anti-Arl8b (56085 S, 1:1000), anti-Atg5 (12994S, 1:1000), anti-Atg7 (2631S, 1:1000) from Cell Signaling Technology; anti-SQSTM1/p62 (ab56416, 1:1000), anti-Tsg101 (ab83, 1:1000), anti-Calreticulin (ab2907, 1:1000) from Abcam; anti-Tim23 (11123-1-AP, 1:1000) from Proteintech, anti-CD63 (PA5-92370, 1:500) from Invitrogen; anti-dendra2 (TA150090, 1:1000) from OriGene; anti-GAPDH (GTX627408, 1:2000) from GeneTex; Anti-Ubiquitin (P4D1) (sc-8017, 1:1000) from Santa Cruz, MTCO1 (459600, 1:1000) from Thermo Fisher Scientific, and MnSOD (06-984, 1:1000) from Millipore. Goat anti-mouse HRP (31430; 1:5000) and Goat-anti-rabbit HRP (31460, 1:5000) secondary antibodies from Thermo Fisher Scientific.

Liang W., Sagar S., Ravindran R., Najor R.H., Quiles J.M., Chi L., Diao R.Y., Woodall B.P., Leon L.J., Zumaya E., Duran J., Cauvi D.M., De Maio A., Adler E.D, & Gustafsson Å.B. (2023). Mitochondria are secreted in extracellular vesicles when lysosomal function is impaired. Nature Communications, 14, 5031.

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