Anti His Monoclonal Antibody

Catalog Number:
LT0426
Lot Number:
411005100202
Isotype:
Mouse IgG2b
Clonality:
HIS.H8
Quantity:
50 μg, Lyophilized with PBS, pH7.4
Positive Control:
Purchase matching Western Blot positive control protein
Immunogen:
HHHHHH (6x His) synthetic peptide conjugated to KLH
Specificity:
Recognizes the epitope of 6x His-tags encoded by many commercially available vectors, regardless of the tag's location in the fusion protein sequences (i.e.: reacts with N-terminal, C-terminal or internal 6x His-tags)
Purification:
Protein A affinity chromatography from mouse ascites fluid
Applications:
Dot, ELISA, IP, IS, WB
Sensitivity:
WB (with ECL): 1-2ng of His-tagged fusion proteins
Working Condition:
WB (with ECL): 1:1000-3000 dilution (incubate for one hour at room temperature)
IS: 1:500-2000 dilution
For best results with other assays (e.g.: Dot, ELISA, IP, etc), please determine optimal working dilution by titration test
Storage:
Store at -20°C for up to 2 years. Reconstitute the lyophilized antibody with deionized water (or equivalent) to a final concentration of 50ug/50ul. Aliquot to avoid repeated freeze-thaw cycles. Store aliquots at 4°C for 1-2 weeks
Images:
anti-His mAb on His-tagged fusion proteins in HEK293 cells
Fig. 01 LEFT: untransfected control; RIGHT: anti-His (in red) on His-tagged fusion proteins in HEK293 cells. Both counterstained with DAPI (in blue)
Comparison between anti-His tag (HIS.H8 / EH158) mAb with 2 different vendor Abs, probed against a standard ladder containing five different His-tagged proteins
Fig. 02 Comparison between anti-His tag (HIS.H8 / EH158) mAb with 2 different vendor Abs, probed against a standard ladder (Qiagen Cat. No. 34705) containing five different His-tagged proteins. All Ab dilutions are 1:2000 (0.5µg/mL)
Standard ladder containing five different His-tagged proteins
Fig. 03 Standard ladder containing five different His-tagged proteins; untransfected control (1), HEK293 cells transfected with His-tagged protein vector (2)
References:

1. Membrane association of monotopic phosphoglycosyl transferase underpins function, Nature Chemical Biology, volume 14, pages538–541 (2018), doi:10.1038/s41589-018-0054-z.

...The primary antibody, mouse anti-His (LifeTein, cat. no. LT0426) was applied in a 1:3,000 dilution for 1?h...

2. Cross-inhibition of ChikunGunya virus fusion and infection by alphavirus E1 domain III proteins, Claudia Sa¡nchez San Martin, Soumya Nanda, Yan Zheng, Whitney Fields and Margaret Kielian, Journal of Virology, Published ahead of print 1 May 2013, doi: 10.1128/JVI.00814-13

... and a mAb against the His Tag (Lifetein LLC, South Plainfield, NJ) to detect His-tagged CHIKV...

3. 14-3-3 Proteins Restrain the Exo1 Nuclease to Prevent Over-resection, The Journal of Biological Chemistry, First Published on April 1, 2015, doi: 10.1074/jbc.M115.644005 jbc.M115.644005.

... ... Anti-FLAG, anti-His (LifeTein, LT0426), anti-14-3-3 (Cell Signaling, 8312), antibodies were purchased from the...

Anti His-tagged proteins
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