GoldiBlot™: Faster His-Tagged Recombinant Protein Detection

Updated: March 13, 2008 Contents See His-tagged Proteins in under an hour Features of GoldiBlot™ Catalog Information Applications See His-tagged Proteins in under an hour Goldiblot™ His Detection Kit is a rapid, sensitive alternative to enzymatic methods for the detection of recombinant His-tagged proteins on membrane blots such as immunoblots and westerns. It uses nitrilotriacetic acid (NTA)-nickel (II)-functionalized gold nanoparticles, which bind tightly and selectively to His tags, followed by metal enhancement for direct visualization.

Goldiblot™ is fast, generating specific magenta bands in one hour with low background, even when the target His-tagged protein is mixed with fifty times as much untagged E coli protein extract. It is also highly sensitive, capable of detecting down to 2.5 ng (example using His-tagged ATF-1 protein). The protocol is simple, involving only one detection agent-binding step. This simplification contributes to the faster speed of the procedure, which can be used to detect quantities of protein in the 25-100 ng range within 45 minutes.

Features of GoldiBlot™

Nickel-NTA Nanogold particles with auto-metallographic amplification provide rapid, sensitive visualization. High sensitivity: low nanogram-level protein detection in one hour. Low background and clean signal. Permanent signal. Western blot detection of His-tagged proteins using GoldiBlot™ His Western Blot kit (total development time: 1 hour). Lane M: All Blue protein ladder. Lanes 1-5: His-tagged ATF-1 loaded at 2.5 – 50 ng (1) 50 ng, (2) 25 ng, (3) 10 ng, (4) 5 ng and (5) 2.5 ng. Lane 6: 100 ng His-tagged YY1. Lane 7: 100 ng His-tagged Src. Lane 8: 50 ng His-tagged Src and bacterial extract with 2,500 ng total E. Coli Protein. Lane 9: bacterial extract with 2,500 ng total E. Coli Protein.

References for GoldiBlot™

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Product and Catalog Information

2090	GoldiBlot™, 15 blots Goldiblot His-tag detection kit, 15 blots. Contains NTA-Ni-Nanogold, metallographic amplification reagent. Product information and instructions (HTML)   Product information (PDF)	$129
2090A	GoldiBlot™, 3 x 15 blots Goldiblot His-tag detection kit, 3 x 15 blots. Contains NTA-Ni-Nanogold, metallographic amplification reagent. Product information and instructions (HTML)   Product information (PDF)	$310

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Applications

• Identify His-tagged proteins rapidly and confidently in cell lysates and extracts.
• Faster, simpler western blots.
• Confirm the expression of his-tagged reporter proteins in transfected cells.

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