Human gapdh endogenous control

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Human GAPDH Endogenous Control is a laboratory reagent that can be used as a reference gene for gene expression analysis. GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) is a commonly used endogenous control gene, as it is consistently expressed across a wide range of cell types and experimental conditions. This product provides a stable and reliable control for normalizing target gene expression data.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

GAPDH (1486 citations) Human GAPDH (26 citations) GAPDH endogenous control (5 citations) Human GAPDH Endogenous Control FAM™ Dye/MGB Probe (2 citations)

6 protocols using human gapdh endogenous control

Quantification of Surface IgM mRNA

Check if the same lab product or an alternative is used in the 5 most similar protocols

IgM+ cells were isolated from WM and HD samples through magnetic cell sorting with anti-human IgM microbeads (Miltenyi Biotec, Auburn, CA, USA) and stored in Trizol at −80 °C. RNA was extracted as per the manufacturer's protocol. complementary DNA was synthesized with the SuperScript VILO kit (Invitrogen Biosource). μ-chain messenger RNA (mRNA) was quantified using the following primer set: 5′- CGTGTCCGAAGAGGAATGG- 3′, 5′-AGAGGCTCAGGAGGAAGAGG-3′, which specifically amplifies surface μ-chain transcripts. Real-time quantitative PCR was performed on QuantStudio 7 flex System (ThermoFisher Scientific) with SYBR Green (Life Technologies) and TaqMan Universal PCR Master Mix (Applied Biosystems). GAPDH was used as a housekeeping control gene (human GAPDH endogenous control, ThermoFisher Scientific). The reaction was performed in duplicate and relative amounts of mRNA transcripts were corrected for the purity of the sample for IgM+ cells and calculated by the comparative ΔCt method using the formula: relative expression=2^ΔCt/ α, where α is the sample's IgM+ cell purity after magnetic sorting. sIgM was measured before magnetic sorting by flow cytometry to correlate mRNA to protein levels.

Argyropoulos K.V., Vogel R., Ziegler C., Altan-Bonnet G., Velardi E., Calafiore M., Dogan A., Arcila M., Patel M., Knapp K., Mallek C., Hunter Z.R., Treon S.P., van den Brink M.R, & Palomba M.L. (2016). Clonal B cells in Waldenström's macroglobulinemia exhibit functional features of chronic active B-cell receptor signaling. Leukemia, 30(5), 1116-1125.

+ Open protocol

+ Expand

Quantification of Gene Expression in Cell Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Gene expression levels of BCL11B, CD45, CD25, CD8α, and CD235α were quantified using TaqMan^® probe and primers (assay IDs: BCL11B Hs01102259_m1 human; CD45 PTPRC Hs 04189704_m1; CD25 IL2RA Hs 00907778_m1; CD8α Hs00233520_m1; CD235α Hs01068072_m1, ThermoFisher Scientific). Human GAPDH endogenous control (assay ID: 4326317E, ThermoFisher Scientific) was used to normalize the data. Real-time PCR was performed in duplicate using the TaqMan^® Fast Advanced Master Mix (ThermoFisher Scientific) using 1.5 μl stock cDNA template. Gene expression data are represented as 2⁻^ΔCt where ΔCt = GAPDH Ct−target gene Ct [17 (link)].

Maskari R.A., Hardege I., Cleary S., Figg N., Li Y., Siew K., Khir A., Yu Y., Liu P., Wilkinson I., O’Shaughnessy K, & Y.a.s.m.i.n. (2018). Functional characterization of common BCL11B gene desert variants suggests a lymphocyte-mediated association of BCL11B with aortic stiffness. European Journal of Human Genetics, 26(11), 1648-1657.

+ Open protocol

+ Expand

Quantitative PCR Analysis of Surfactant Protein Genes

Check if the same lab product or an alternative is used in the 5 most similar protocols

The TaqMan® Gene Expression Master Mix was used together with the TaqMan® Gene Expression Assays SFTPA1: Hs01921510_s1, SFTPA2: Hs00359837_m1, SFTPB: Hs00167036_m1, SFTPC: Hs00161628_m1, and SFTPD: Hs00358340_m1 (Applied Biosystems, Foster City, CA, USA). 100 ng of cDNA and Human GAPDH Endogenous Control (Applied Biosystems, Foster City, CA, USA) were added to each reaction. The choice of GAPDH as a reference gene was based on GAPDH having a stringent correlation in the lung cells and the least variation of the seven common reference genes [21 (link)]. The PCR was performed in a 96-well plates format (25 μl per well) on a 7500 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The temperature settings were as follows: step 1 at 50°C for 2 minutes, step 2 at 95°C for 10 minutes, step 3 at 95°C for 15 seconds, and step 4 at 60°C for 1 minute. Steps 3 and 4 were repeated 40×. The expression data were calculated using the comparative ΔCt method as described elsewhere [22 (link)]. Only normalization to the housekeeping gene (GAPDH) was used.

Ručka Z., Koutná I., Tesařová L., Potěšilová M., Stejskal S., Šimara P., Vaňhara P., Doležel J., Zvoníček V., Coufal O, & Čapov I. (2014). Intravenous insulin therapy during lung resection does not affect lung function or surfactant proteins. BMC Pulmonary Medicine, 14, 155.

+ Open protocol

+ Expand

Quantifying CADM1 Expression in Cancer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated from parental T24, T24-Ad-CADM1, T24-Ad-sh1/2/3 and T24-Ad-GFP1/2 cells using the TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. qPCR was performed on ABI 7900 96 HT series PCR machine (Applied Biosystems; Thermo Fisher Scientific, Inc.) using FastStart Universal SYBR Green Master (ROX) kit (Roche). qPCR was then performed with the specific CADM1 primers (forward, 5′-ATGGCGAGTGTAGTGCTGC-3′ and reverse, 5′-GATCACTGTCACGTCTTTCGT-3′). Relative CADM1 expression level was then normalized to levels of GAPDH (forward, 5′-GGAGCGAGATCCCTCCAAAAT-3′ and reverse, 5′-GGCTGTTGTCATACTTCTCATGG-3′) as an internal control. PCR conditions were 50°C for 2 min and 95°C for 10 min followed by 40 cycles of 95°C for 15 sec, and 60°C for 1 min. The expression levels were normalized against glyceraldehyde 3-phosphatedehydrogenase (GAPDH) (Human GAPDH Endogenous Control, Applied Biosystems; Thermo Fisher Scientific, Inc.). The results were summarized from triplicate experiments using the 2^−∆∆Cq method (30 (link)).

Chen Y., Liu L., Guo Z., Wang Y., Yang Y, & Liu X. (2018). Lost expression of cell adhesion molecule 1 is associated with bladder cancer progression and recurrence and its overexpression inhibited tumor cell malignant behaviors. Oncology Letters, 17(2), 2047-2056.

+ Open protocol

+ Expand

Luteolin Modulates Inflammatory Response

Check if the same lab product or an alternative is used in the 5 most similar protocols

Keratinocytes were pretreated with luteolin (10–100 µM, 30 min) before TNF stimulation (50 ng/mL, 6 h). Total RNA was extracted with an RNeasy Mini kit (Qiagen Inc., Valencia, CA). For human skin samples, total RNA was extracted using a Fibrous Tissue mini kit (Qiagen). An iScript cDNA synthesis kit (BioRad, Hercules, CA) was used for reverse-transcription of each sample. qRT-PCR was performed using Taqman gene expression assays (Applied Biosystems, Foster City, CA) for IL-6, IL-8, VEGF, NFKB1 and RELA. Samples were run using a 7300 Sequence Detector, according to TaqMan Gene Expression Assay instructions (Applied Biosystems). The mRNA expression was determined from standard curves run with each experiment. Relative mRNA levels were normalized to human GAPDH endogenous control (Applied Biosystems).

Weng Z., Patel A.B., Vasiadi M., Therianou A, & Theoharides T.C. (2014). Luteolin Inhibits Human Keratinocyte Activation and Decreases NF-κB Induction That Is Increased in Psoriatic Skin. PLoS ONE, 9(2), e90739.

+ Open protocol

+ Expand

Quantitative PCR Analysis of Runx2

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated using a Mag Extractor (Toyobo, Osaka, Japan) according to the manufacturer's protocol, and single-stranded cDNA was synthesized using a High-Capacity RNA-to-cDNA Master Mix (Applied Biosystems, Foster City, CA, USA). Runx2 mRNA levels were analyzed by quantitative real-time PCR using a TaqMan ® Gene Expression Assay (Hs00231692 mL; Applied Biosystems) on a Step One Plus PCR system (Applied Biosystems). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was co-amplified as an internal standard (Human GAPDH endogenous control; Applied Biosystems). Gene expression was measured using the ddCt method 12) .

Hashimoto Y., Nishikawa H., Kusunoki M., Li P, & Hontsu S. (2015). A novel membrane-type apatite scaffold engineered by pulsed laser ablation. Dental materials journal, 34(3).

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!