Description
Key facts
Applications:IHC-PSee reactivity dataSee the reactivity data table below for information on validated species and application combinations.,
Target:Antibody DiluentSee target data,
Form:LiquidSee storage information,
Storage buffer:pH: 7 - 7.8Preservative: 0.05% Sodium azide
Product details:
Antibody Diluent ab64211 is used to dilute primary antibodies to their working concentrations for use in IHC. It can also be used for convenient storage of diluted primary antibodies.
The antibody diluent is formulated to reduce background and non-specific binding of the primary antibody. In most cases, antibodies can be stored in the diluent, at their working concentrations for IHC, for up to 24 months at 2-8°C.
The diluent is based on a Tris buffer, and contains a stabilizing carrier protein, a wetting agent to reduce surface tension, an anti-microbial agent, and a green dye. It should not be used with HRP-conjugated primary antibodies.
IHC protocol suitable for use with Antibody Diluent ab64211:
For fluorescent IHC, skip step 3, incubate with fluorescent dye conjugated secondary at step 6, skip rest of steps, and mount with
anti-fade mounting medium
1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.
2. Use appropriate
antigen retrieval buffer or enzyme
(primary antibody dependent) to treat sections. Wash 3 times in buffer.
3. Add enough
hydrogen peroxide blocking solution
to cover the sections. Incubate for 10 minutes. Wash 2 times in buffer. If necessary, use
avidin biotin blocking
4. Apply
protein block
normal serum
from same species as secondary antibody) and incubate for 5 minutes at room temperature to block nonspecific background staining. Wash once in buffer.
Apply primary antibody in Antibody Diluent ab64211 and incubate.
6. Wash 4 times in buffer. Incubate slide with
biotinylated secondary antibody
HRP polymer secondary antibody
and skip step 7). Wash 4 times in buffer.
7. Apply
streptavidin-HRP
and incubate for 10 minutes at room temperature.
8. Rinse 4 times in buffer. Place slide in
DAB substrate
AEC substrate
and incubate until desired color is achieved (1-10 mins). Rinse 4 times in buffer.
9. Add enough drops of
hematoxylin
to cover the section. Incubate for 1 minute.
10. Rinse 7-8 times in tap water. Add
mounting medium
to cover the section.
Find complete IHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the
IHC kits and reagents guide
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C
