Application of His-tagged antibodies in WB protein screening and ELISA analysis

The His tag is an additive to a popular recombinant protein to facilitate the purification and detection of proteins

Figure 4.

Figure 6
.
Rabbit Anti-His Tag (300-005-240) Indirect Sandwich ELISA Method for Detection of His-Tagged Proteins
.

1.
Capture antibody: AffiniPure Rabbit Anti-His Tag (300-005-240) is coated with 1 μg/well (100 μl, 10 μg/ml).

Block the plate
in PBST with 1% Bovine Serum Albumin (IgG-Free, Protease-Free) (001-000-162) 2.
Sample: His-Tagged SARS-CoV-2 Spike Protein RBD (Genscript - Z03483-100), loaded at 20 ng/well (100 μl @ 200 ng/ml
。
3.
Detection antibody 1: human antisars -cov-2 IgG Spike S1 antibody (Genscript A02038
).
First hole 40 ng (100 ul @ 400 ng/ml); Serial dilution 1:2 across plates
.

4.
Detection of antibody 2: HRP Goat Anti-Human IgG, Fcγfragment specific (min X Bov, Ms, Rb Sr Prot) (109-035-170) dilution 1:20 000 (100 μl/well).
5.
Plate incubate with TMB for 30 min at room temperature and read at 620 nm with a plate reader
.

*Experimental tip: When using rabbit antibodies during capture, it is recommended to use detection antibodies that are cross-adsorbed with rabbit serum proteins to avoid background generation
.
Jackson Immuno Research's antibodies, including anti-human antibodies, can eliminate cross-reactivity to a variety of hosts, allowing different host species to eliminate high background
.