Amplite® West ECL HRP Substrate Femto Sensitivity

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Fax	1-800-609-2943
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H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22

Overview SDS Protocol

Amplite® West ECL HRP Substrate is an ultra-sensitive chemiluminescent (ECL) substrate formulated for highest sensitivity and stability. It is used for detection of peroxidase activity from HRP-conjugated antibodies and probes on nitrocellulose or PVDF membranes. The substrate provides a quick and efficient way for detecting proteins expressed at femtogram levels. It does not require any expensive instruments and consumables and is compatible with the existing instruments commonly used for western blots. When combined with very diluted and optimized antibody concentrations and blocking buffers, Amplite® West ECL HRP Substrate enables detection of target proteins in small amounts.

Components

Example protocol

AT A GLANCE

Protocol Summary

Prepare working solution by mixing equal parts of Solution A and Solution B
Incubate membrane with working solution
Image with an imaging system or X-ray film

SAMPLE EXPERIMENTAL PROTOCOL

After incubating with the HRP enzyme conjugated antibody, wash the membrane 6 times for 5 minutes each inproper wash buffer to remove any unbounded conjugates.
Prepare the developing working solution by mixing equal parts of Solution A and Solution B. Note: Use a sufficient volume to ensure the blot is completely wetted and will not dry out during development (0.1 mL/cm²)
Incubate the membrane with the working solution for 5 minutes.
Remove membrane from the working solution and wash briefly with wash buffer, remove excess liquid with an absorbent tissue if necessary.
Image the blot using an imaging system or X-ray film.

References

View all 50 references: Citation Explorer

Self-luminescent photodynamic therapy and pathogen detection for infectious diseases.
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Journal: Mikrochimica acta (2021): 33

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Authors: Wang, Di and Li, Danting and Zhang, Yuxin and Chen, Jie and Zhang, Ying and Liao, Chuyao and Qin, Siyuan and Tian, Yuan and Zhang, Zunjian and Xu, Fengguo
Journal: Acta pharmaceutica Sinica. B (2021): 763-780

Human platelets display dysregulated sepsis-associated autophagy, induced by altered LC3 protein-protein interaction of the Vici-protein EPG5.
Authors: Schwertz, Hansjörg and Rowley, Jesse W and Portier, Irina and Middleton, Elizabeth A and Tolley, Neal D and Campbell, Robert A and Eustes, Alicia S and Chen, Karin and Rondina, Matthew T
Journal: Autophagy (2021): 1-17

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Journal: Analytica chimica acta (2021): 83-90

Analysis of Proteins by Immunoblotting.
Authors: Kielkopf, Clara L and Bauer, William and Urbatsch, Ina L
Journal: Cold Spring Harbor protocols (2021): pdb.prot102251

Raft-like lipid microdomains drive autophagy initiation via AMBRA1-ERLIN1 molecular association within MAMs.
Authors: Manganelli, Valeria and Matarrese, Paola and Antonioli, Manuela and Gambardella, Lucrezia and Vescovo, Tiziana and Gretzmeier, Christine and Longo, Agostina and Capozzi, Antonella and Recalchi, Serena and Riitano, Gloria and Misasi, Roberta and Dengjel, Joern and Malorni, Walter and Fimia, Gian Maria and Sorice, Maurizio and Garofalo, Tina
Journal: Autophagy (2021): 2528-2548

Platelet autophagic machinery involved in thrombosis through a novel linkage of AMPK-MTOR to sphingolipid metabolism.
Authors: Lee, Tzu-Yin and Lu, Wan-Jung and Changou, Chun A and Hsiung, Yuan-Chin and Trang, Nguyen T T and Lee, Cheng-Yang and Chang, Tzu-Hao and Jayakumar, Thanasekaran and Hsieh, Cheng-Ying and Yang, Chih-Hao and Chang, Chao-Chien and Chen, Ray-Jade and Sheu, Joen-Rong and Lin, Kuan-Hung
Journal: Autophagy (2021): 4141-4158

Fe-MOGs-based enzyme mimetic and its mediated electrochemiluminescence for in situ detection of H2O2 released from Hela cells.
Authors: Zong, Li-Ping and Ruan, Ling-Yu and Li, Junji and Marks, Robert S and Wang, Jun-Song and Cosnier, Serge and Zhang, Xue-Ji and Shan, Dan
Journal: Biosensors & bioelectronics (2021): 113216

Essential meiotic structure-specific endonuclease1 (EME1) promotes malignant features in gastric cancer cells via the Akt/GSK3B/CCND1 pathway.
Authors: Guo, Zhiguo and Liang, Erbo and Li, Wei and Jiang, Leilei and Zhi, Fachao
Journal: Bioengineered (2021): 9869-9884