Hrp conjugated gapdh antibody

Manufactured by Proteintech

Sourced in China

The HRP conjugated GAPDH antibody is a laboratory reagent designed for the detection of the GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) protein in biological samples. GAPDH is a common housekeeping gene used as a loading control in various immunoassays. The HRP (Horseradish Peroxidase) conjugation allows for the direct detection of GAPDH through a colorimetric or chemiluminescent signal.

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GAPDH (1973 citations) GAPDH antibody (119 citations) GAPDH-HRP (13 citations) HRP-conjugated GAPDH monoclonal antibody (6 citations) HRP-conjugated GAPDH (4 citations) HRP-conjugated GAPDH Monoclonal antibody (HRP-60004 (2 citations)

3 protocols using hrp conjugated gapdh antibody

Western Blot Analysis of Yap Proteins

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Yap protein levels were determined by western immunoblotting analysis. In brief, proteins isolated from cells (40 μg) were separated by electrophoresis on a 10% SDS polyacrylamide gel. Partitioned proteins were transferred to a nitrocellulose membrane. The membrane was probed with rabbit anti-Yap antibody (Cell Signaling Technology), rabbit anti-PABP antibody (Abcam), rabbit anti-eIF4G antibody (Cell Signaling Technology), mouse anti-eIF4A, anti-eIF4B or anti-eIF4E (Santa Cruz Biotechnology). HRP-conjugated anti-rabbit or anti-mouse IgG antibody (Jackson ImmunoResearch) was used as the secondary antibody. The corresponding protein bands were visualized using enhanced chemiluminescence reagents. The same membranes were re-probed with HRP conjugated GAPDH antibody (Proteintech), mouse anti-alpha tubulin antibody (Santa Cruz Biotechnology) or mouse anti-PCNA (Santa Cruz Biotechnology) to confirm equal loading of proteins for each sample.

Wu N., Yuan Z., Du K.Y., Fang L., Lyu J., Zhang C., He A., Eshaghi E., Zeng K., Ma J., Du W.W, & Yang B.B. (2019). Translation of yes-associated protein (YAP) was antagonized by its circular RNA via suppressing the assembly of the translation initiation machinery. Cell Death and Differentiation, 26(12), 2758-2773.

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Immunoblotting Analysis of Protein Expression

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Immunoblotting was performed as previously described [20 (link)]. After gradient separated by SDS-PAGE and blotted to polyvinylidene fluoride membranes, the expressing levels of target proteins in arteria or A7r5 cells were assessed using a chemiluminescence system with an enhanced chemiluminescence (ECL) reagent (Thermo Scientific, USA). The following primary antibodies were used: phosphor-Akt (Thr308) (CST, Beverly, MA, USA) and β-catenin (CST, Beverly, MA, USA). The HRP-conjugated GAPDH antibody (Proteintech, Wuhan, China) served as an internal standard for normalization. The densitometric analysis was performed using Image Lab 4.1 (Bio-Rad, USA).

Zhou J., Zhang C., Zheng G.H, & Qiu Z. (2018). Emblic Leafflower (Phyllanthus emblica L.) Fruits Ameliorate Vascular Smooth Muscle Cell Dysfunction in Hyperglycemia: An Underlying Mechanism Involved in Ellagitannin Metabolite Urolithin A. Evidence-based Complementary and Alternative Medicine : eCAM, 2018, 8478943.

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Western Blot Protein Detection Protocol

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The expression of all ESFs were detected by Western blotting. The total cell pellets were completely lysed in 1×SDS-PAGE loading buffer (mixture of 2×Laemmli Sample Buffer (#1610737, Bio-Rad), RIPA buffer (#89900, Thermo Fisher) and Protease Inhibitor (#04693116001, Roche)) and heated at 99°C for 10 mi n. The samples were subsequently separated by SDS-PAGE gels and transferred to PVDF membranes. The following antibodies were then used for detection: Monoclonal ANTI-FLAG M2 antibody (F1084, Sigma-Aldrich), IRDye 800CW Goat anti-Mouse IgG (#926–32210, LI-COR), Anti-mouse IgG antibody (#7076, Cell Signaling Technology), GAPDH antibody (FL-335) (sc-25778, Santa Cruz Biotechnology), IRDye 680RD Goat anti-Rabbit IgG (#926–68071, LI-COR) and goat anti-rabbit IgG-HRP antibody (sc-2004, Santa Cruz Biotechnology), GAPDH (14C10) Rabbit mAb (#2118S, Cell Signaling Technology), Anti-rabbit IgG, HRP-linked Antibody (#7074S, Cell Signaling Technology), HRP-Conjugated GAPDH Antibody (#HRP-60004, Proteintech). The fluorescence labeling second antibodies were detected and visualized by Odyssey Imaging Systems (LI-COR) directly. The HRP conjugated primary or second antibodies were detected using an enhanced chemiluminescence detection kit and visualized by ChemiDoc Touch Imaging System (Image Lab, Bio-Rad).

Mao M., Hu Y., Yang Y., Qian Y., Wei H., Fan W., Yang Y., Li X, & Wang Z. (2018). Modeling and predicting the activities of trans-acting splicing factors with machine learning. Cell systems, 7(5), 510-520.e4.

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