The expression of mRNA was semi-quantified using RT-PCR with the TaqMan® Gene Expression Assays, Step One Plus Real-Time PCR system, and Step One Software (Life Technologies). After culturing cells at 80% confluence in 6-well culture plates, the total RNA was extracted using the RNeasy® Mini kit (cat. no. 74104; Qiagen, Venlo, Limburg, The Netherlands) and the cDNA was immediately synthesized using SuperScript® First-Strand Synthesis for RT-PCR (cat. no. 11904-018; Life Technologies) for RT-PCR according to the manufacturer's instructions. The gene expression data were calculated relative to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) level. TaqMan probes for GAPDH, E-cadherin, vimentin, fibronectin, slug, and PD-L1 were obtained by ordering from Applied Biosystems through Life Technologies (assay identification numbers: Hs02758991_g1, Hs01023894_m1, Hs00185584_m1, Hs00365052_m1, Hs00950344_m1 and Hs01125301_m1).
Taqman probes for gapdh
Manufactured by Thermo Fisher Scientific
Sourced in United States
TaqMan probes for GAPDH are molecular biology reagents used in real-time PCR assays to detect and quantify the expression of the GAPDH gene, which is a commonly used reference gene. The probes are designed to specifically hybridize to the GAPDH sequence and emit a fluorescent signal during the PCR amplification process, enabling the measurement of GAPDH levels in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions)
Stepone software, by Thermo Fisher Scientific (1 mentions)
Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions)
Primescript rt reagent, by Takara Bio (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
7500 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Rnu48, by Thermo Fisher Scientific (1 mentions)
Antibodies against gapdh, by Santa Cruz Biotechnology (1 mentions)
Trametinib, by Selleck Chemicals (1 mentions)
Mir 145 5p, by Thermo Fisher Scientific (1 mentions)
Mir 204 5p, by Thermo Fisher Scientific (1 mentions)
Mir 199b 5p, by Thermo Fisher Scientific (1 mentions)
Vemurafenib, by Selleck Chemicals (1 mentions)
Nutristem hpsc xf medium, by Sartorius (1 mentions)
Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Stepone real time pcr system, by Thermo Fisher Scientific (1 mentions)
Taqman fast advanced master mix, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Rnase free dnase, by Qiagen (1 mentions)
4 protocols using taqman probes for gapdh
1
Real-Time PCR for Epithelial-Mesenchymal Transition
2
Quantitative RT-PCR Analysis of VASH2 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from cells and tissues using TRIzol (Invitrogen, 15596–026, Carlsbad, CA, USA), and cDNA was synthesized using the Primescript RT Reagent (TAKARA, DRR037A, Dalian, China). Quantitative RT-PCR was performed on a 7500 Real-Time PCR System (Applied Biosystems) using Taqman probes for GAPDH (Hs99999-m1, Applied Biosystems) and VASH2 (Hs00226928-m1). GAPDH was used as a reference to obtain the relative fold change for targets using the comparative Ct method.
3
Molecular Targets in Cancer Biology
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies against GAPDH, Bcl-2 and VEGF were obtained from Santa Cruz Biotechnology. Vemurafenib and trametinib were obtained from Selleck Chemicals. TaqMan probes for GAPDH, VEGFA, TIMP2, PTPN14, HIF-1α, Bcl-2, CCL3, IL1b, CCL5, tgfB, CXCL8, CSF3, PDGFb, CCL20, miR-4443, miR-4488, miR-204-5p, miR-199b-5p, miR-630, miR-1234, ts-3676, miR-145-5p and RNU48 were purchased from Applied Biosystems. Primers relative to H3, RAD51, Notch1 and FOXM1 have been used in the work by Canu et al [18 (link)]. Western Blot analysis were performed as previously described [11 (link), 16 (link)].
4
qPCR Analysis of Ovarian Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was isolated using the RNeasy Mini Kit and treated with RNase-free Dnase (both from Qiagen, Germany). For cDNA synthesis, 10 ng of total RNA was reversely transcribed using random hexamers and Superscript III reverse transcriptase (Invitrogen, Thermo Fisher Scientific). Samples were subjected to qPCR using TaqMan probes for GAPDH (Hs02786624_g1), MCAM (Hs00174838_m1), and TAGLN (Hs01038777_g1) and TaqMan Fast Advanced Master Mix (all from Thermo Fisher Scientific) on the StepOne real-time PCR System (Applied Biosystems, USA). As negative control, the human embryonic stem cell line HS980 was included after culture in NutriStem hPSC XF medium (Biological Industries, Israel) on hrLN-521-coated plates (Biolamina, Sweden)65 (link). The experiment was performed on three undissociated pieces of whole ovary tissue from two donors, respectively, three independently dissociated ovarian tissue samples, and three independently sorted DDX4 Ab+ and Ab− cell fractions. All samples were run in technical duplicates. Results are presented as mean ± SD (standard deviation of mean).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!