Enhanced chemiluminescense detection kit

Manufactured by Merck Group

Sourced in United States

The Enhanced Chemiluminescence Detection Kit is a laboratory product designed to facilitate the detection and visualization of proteins and other biomolecules in Western blot analyses. The kit utilizes a chemiluminescent reaction to produce a light signal, which can then be captured and analyzed using appropriate imaging equipment.

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Lab products found in correlation

Pvdf membrane, by Merck Group (1 mentions) Gapdh mb001, by Bioworld Technology (1 mentions) Ha 11 mms 101p, by Fortrea (1 mentions) Ripa buffer, by Beyotime (1 mentions) Hrp conjugated goat anti rabbit igg, by EarthOx (1 mentions)

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Chemiluminescense (2 citations)

2 protocols using enhanced chemiluminescense detection kit

Western Blot Analysis of Protein Expression

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Cells were washed with PBS and lysed with RIPA buffer supplemented with 1.0 mM PMSF. After 15-min incubation on ice, lysates were centrifuged at 12,000 rpm for 20 min at 4 °C. The supernatant was collected, and the protein concentration was measured by bicinchoninic acid (BCA) assay (Beyotime, China). Whole cell lysates (25–50 μg of denatured protein) were separated on a 4–20 % gradient SDS polyacrylamide gel and then transferred to a PVDF membrane (Millipore). The membrane was blocked for 1 h at room temperature in PBST, pH 7.4 (PBS with 0.1 % Tween-20) with 5 % BSA. After washing, the membrane was incubated with primary antibodies overnight at 4 °C, followed by extensive washing and incubation with HRP-conjugated secondary antibodies. At last, protein was visualized by an enhanced chemiluminescense detection kit (Millipore, USA). After stripping, the blot was reprobed with β-Actin or GAPDH antibody (Santa Cruz) to ensure equal loading of total protein.

Li X., Zhang G., Chen Q., Lin Y., Li J., Ruan Q., Chen Y., Yu G, & Wan X. (2016). CD317 Promotes the survival of cancer cells through apoptosis-inducing factor. Journal of Experimental & Clinical Cancer Research : CR, 35, 117.

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Protein Expression Analysis Protocol

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Whole-cell lysate was prepared by suspending cells in the RIPA buffer (Beyotime, China) supplemented with 1× complete protease inhibitors mixture and 1× phosphatase inhibitor (Roche). Protein concentration was determined by BCA assay (Pierce, Rockford, USA). Equal quantities of proteins were separated by SDS/PAGE, transferred to a PVDF membrane, and blotted with specific antibodies. Protein in the membrane was visualized by an enhanced chemiluminescense detection kit (Millipore, USA). Rabbit antibodies against the following proteins or modifications were used with the catalogue numbers and sources indicated: CD317 (ab134061), TGF-α (ab208156) (Abcam); caspase3 (9662) (CST); pY1068 EGFR (BS5010), pY845 EGFR (BS5013), pY705 STAT3 (BS4181), STAT3 (AP0365), ERK½ (BS1112), pT202/Y204 ERK½ (BS5016), cyclin D1 (BS6352), and p16 INK4a (BS6431), caveolin-1 (BS9878M) (Bioworld Technology), and AREG (16036–1-AP) (Proteintech). Mouse antibodies against the following proteins/epitopes were used: HA.11 (MMS-101P) (Covance); Ki67 (P6834) (Sigma); transferrin receptor (13–6800) (Invitrogen); GAPDH (MB001) (Bioworld Technology); β-actin (sc-47778) and EGFR (sc-373746) (Santa Cruz Biotech). HRP-conjugated mouse anti-His (M20020) was purchase from Abmart., mouse HRP-conjugated goat anti-mouse IgG (074–1806) from KPL, and HRP-conjugated goat anti-rabbit IgG (E030120–02) from EARTHOX.

Zhang G., Li X., Chen Q., Li J., Ruan Q., Chen Y.H., Yang X, & Wan X. (2019). CD317 activates EGFR by regulating its association with lipid rafts. Cancer research, 79(9), 2220-2231.

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