Production and characterization of polyclonal antibody against Bengal tiger (Panthera tigris tigris) immunoglobulin G

Figures & data

Figure 1. Analysis of orange-haired Bengal tiger IgG. (A) The purified orange-haired Bengal tiger IgG was analyzed under reducing and non-reducing conditions using SDS-PAGE and stained with Coomassie Brilliant Blue. Lanes M: molecular mass standards; 1: Non-reducing conditions; 2: Reducing conditions. (B) The non-purified and purified rabbit anti-Bengal tiger IgG polyclonal antibody were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. Lanes M: molecular mass standards; 1–2: Non-purified rabbit anti-tiger IgG; 3–4: Purified rabbit anti-tiger IgG.

Figure 2. Evaluation of the binding capacity of rabbit anti-Bengal tiger IgG polyclonal antibody to Bengal tiger IgG. The reactive band was observed at approximately 170 kDa.

Figure 3. Determinations of optimal dilution of indirect ELISA reagents as presented by mean OD values at different coating antigen concentration of 0.5 µg/ml (A) 1 µg/ml (B) 2 µg/ml (C).

Note: ^aOrange-haired Bengal tiger IgG. ^bHRP-conjugated goat anti-rabbit IgG. ^cRabbit anti-Bengal tiger IgG polyclonal antibody generated in this study. ‘*’ indicates the suitable OD value from this study.

Table 1. Species specificity of rabbit anti-Bengal tiger immunoglobulin G and cross-reactivity among wild felid species (family Felidae).

Animal	OD	%
Bengal tiger: orange hair^a,b	1.194	100.00
Bengal tiger: white hair^a,b	0.903	75.63
Golden tabby tiger^a,b	0.893	74.79
Snow-white tiger^a,b	0.849	71.11
Siberian tiger^a,b	0.868	72.70
Lion^b	0.641	53.69
Lion: white hair^b	0.692	57.96
Panther^b	0.813	68.10
Leopard^b	0.794	66.50
Jaguar^b	0.892	74.71
Clouded leopard^b	0.844	70.69
Asian golden cat	0.776	64.99
Leopard cat	0.815	68.26
Jungle cat	0.851	71.27

^a Panthera tigris subspecies.

^bLocated in the Panthera genus.

Figure 4. The immunoblotting analysis of rabbit anti-Bengal tiger IgG polyclonal antibody cross-reactivity with domestic animals. The cross-reactivity was strong with the cat IgG and faint with the pig IgG.

Table 2. Cross-reactivity of purified rabbit anti-Bengal tiger immunoglobulin G with domestic animals’ immunoglobulin G.

Animal	OD	%
Bengal tiger	0.593	100.00
Domestic cat*	0.377	63.58
Pig*	0.228	38.45
Cow	0.062	10.46
Sheep	0.043	7.25
Goat	0.051	8.60
Chicken	0.041	6.91

‘*’ indicates that the cross-reactivity was observed in this study by the rabbit anti-Bengal tiger IgG polyclonal antibody reactivity with domestic animals’ IgG.