Bio-Rad Immun-Blot® Opti-4CN™ Colorimetric Kits - Manual (Page 4)

170-8239 Amplified Opti-4CN Detection Kit, Goat-anti-Rabbit
All components of 170-8238
Goat-anti-Rabbit-HRP conjugated secondary
antibody, 0.5 ml
170-8240 Amplified Opti-4CN Detection Kit, Goat-anti-Mouse
All components of 170-8238
Goat-anti-Mouse-HRP conjugated secondary
antibody, 0.5 ml
Additional required items not provided in these kits
All kits Nitrocelluose or PVDF membrane
Primary antibody
Tween-20, 10 ml, sufficient for 2,500 cm
2
Bovine serum albumin, 7.5 g, sufficient for
2,500 cm
2
170-8235 Blocker
Buffer
HRP-linked secondary antibody
170-8236/7 Blocker
Buffer
170-8238 Dimethyl sulfoxide (DMSO), 500 ml, sufficient
for 2,500 cm
2
HRP-linked secondary antibody
170-8239/40 Dimethyl sulfoxide (DMSO), 500 ml, sufficient
for 2,500 cm
2
1.4 Product Storage and Stability
The kit is shipped at 4 °C. Store the unopened kit
at 4°C. Powdered blocker, powdered PBS and PBS solu-
tions may be stored at room temperature; all other com-
ponents are stored at 4 °C. After being put into solution,
the blocker should be stored at 4 °C. All kit compo-
nents are guaranteed for 1 year from the time of receipt.
The membrane with bound antigen is then incubated
with a primary antibody specific to the antigen of interest.
The blot is washed to remove unbound antibody, incubat-
ed with a secondary antibody linked to HRP, and then
washed again to remove unbound secondary antibody. If
there is no amplification involved (i.e., catalog numbers
170-8235 through 170-8237), then the blot is incubated in
the Opti-4CN substrate for up to 30 minutes, or until the
desired sensitivity is attained. If the signal is to be amplified,
the blot is incubated in the Bio-Rad amplification reagent
(BAR), washed, incubated in streptavidin-HRP, and washed
again before being incubated in the Opti-4CN substrate.
1.3 Kit Components
170-8235 Opti-4CN Substrate Kit
Opti-4CN substrate, 12.5 ml
Opti-4CN diluent concentrate, 10x, 62.5 ml
170-8236 Opti-4CN Detection Kit, Goat-anti-Rabbit
Opti-4CN substrate, 12.5 ml
Opti-4CN diluent concentrate, 10x, 62.5 ml
Goat-anti-Rabbit-HRP conjugated secondary
antibody, 0.5 ml
170-8237 Opti-4CN Detection Kit, Goat-anti-Mouse
Opti-4CN substrate, 12.5 ml
Opti-4CN diluent concentrate, 10x, 62.5 ml
Goat-anti-Mouse-HRP conjugated secondary
antibody, 0.5 ml
170-8238 Amplified Opti-4CN Substrate Kit
Bio-Rad Amplification Reagent, 53 ml
Streptavidin-HRP, 0.5 ml
Blocker, 20 g
PBS, powder to make 1 liter of 10x PBS
2x Amplification diluent, 106 ml
Opti-4CN substrate, 12.5 ml
Opti-4CN diluent concentrate, 10x, 62.5 ml
2 3
4100130c.qxd 4/9/99 8:29 AM Page 2