Excellent His-Tag Antibody for Western Blot and ELISA

February 24, 2016

Biomedical Engineering
Boston University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

GenScript USA, Inc.

Product Name:

THE(TM) His Tag Antibody, mAB, mouse

Catalog Number:

A00186-100

I used this product to detect low levels of His-tagged protein (induced synthesis) in bacterial crude lysate via Western Blot, and as a primary antibody in an indirect ELISA. It was extremely sensitive and highly specific in both assays, at the manufacturer's recommended concentration, 0.1 ug/mL.

Experimental Design and Results Summary

Applications

Western Blot

Sample

His-tagged Camelid nanobody proteins (17.6 kd) in crude bacterial cell lysate

Primary Incubation

NA

Blocking Agent

Genscript 's ONE-HOUR Western kit, Pre-block A+B, 5 minutes at Room Temp

Secondary Incubation

Genscript 's ONE-HOUR Western kit, THE His Tag Antibody in buffer WB-1 & WB-2 (0.1 ug/mL), 40 minutes, Room Temp

Tertiary Incubation

TMB substrate, ~2 min then wash with dd H20 & dry in dark

Detection

Versadoc light imaging

Results Summary

This antibody was highly sensitive, and strongly detected the His-tagged target proteins, although the band did not show up with coomassie staining. It was also highly specific and did not strongly label two large bands of non-His tagged protein.

Additional Notes

None

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Summary

The Good

Highly sensitive and specific. Stores very well - I thawed an aliquot that was 1.5 years old (stored at -22 C), and it worked as well as brand new.

The Bad

None

The Bottom Line

Great antibody for detecting HIS tagged proteins (6X His tag) in Western or ELISA.

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