Pan Cytokeratin Antibody – Mouse Monoclonal

Monoclonal

IgG1

AE1/AE3

Cytokeratin AE1+ AE3 antibody was produced in Mouse.

Bovine, Chicken, Human, Monkey, Mouse, Rabbit, Rat

10mM PBS, pH 7.4, with 0.2% BSA and 0.09% Sodium Azide

Protein G Purified

+4°C, avoid freezing

Cytokeratin AE1+AE3

Human epidermal keratin.

Purified protein

Cytokeratin AE1+AE3 antibody was raised against human epidermal keratin.

Due to the highly specific nature of antibodies and antigens, we cannot predict or be held responsible with respect to how this antibody will behave in your systems. Researchers using this antibody should conduct optimization studies to achieve the most optimal result possible for their intended application.

The following protocol is a recommendation only, and MyBioSource, Inc. makes no guarantee of the results:

Tissue Preparation:
Formalin fixation and embedding in paraffin wax.

Tissue Sectioning:
Make 4-um sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-dryingoven for 45 minutes at 60°C.

Deparaffinization:
Wash dry slides in 3 changes of xylene - 5 minutes each @ RT

Rehydration:
Wash slides in 3 changes of 100% alcohol - 3 minutes each @ RT
Wash slides in 2 changes of 95% alcohol - 3 minutes each @ RT
Wash slides in 1 change of 80% alcohol - 3 minutes @ RT
Rinse slides in gentle running distilled water - 5 minutes @ RT

Antigen retrieval:
Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes
Remove from heat and let stand at room temperature in buffer - 20 minutes
Rinse in 1X TBS with Tween (TBST) -1 minute @ RT

Immunostaining:
(Do not allow tissues to dry at any time during the staining procedure)
Apply a universal protein block - 20 minutes @ RT
Drain protein block from slides, apply diluted primary antibody - 45 minutes @ RT
Rinse slides in 1 X TBST - 1 minute @ RT
Apply a biotinylated secondary antibody appropriate for the primary antibody - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase streptavidin - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase chromogen substrate - 30 minutes @ RT
Wash slides in distilled water - 1 minute @ RT

Dehydrate:
(This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB)
Wash slides in 2 changes of 80% alcohol - 1 minute each @ RT
Wash slides in 2 changes of 95% alcohol - 1 minute each @ RT
Wash slides in 3 changes of 100% alcohol - 1 minute each @ RT
Wash slides in 3 changes of xylene - 1 minute each @ RT
Apply coverslip

During shipment,small volumes of product will ocassionally become entrapped in the seal of the product vial. We recommend briefly centrifuging the vial to dislodge any liquid in the container's cap prior to opening.

This reagent may contain sodium azide. The chemical, physical, and toxicologial properties of this material have not been thoughly investigated.Standard Laboratory Practices should be followed. Avoid skin and eye contact,inhalation, and ingestion. Sodium azide forms hydrazoic acid under acidic conditions and may react with lead or copper plubing to form highly explosive metal azides. On disposal, flush with large volumes of water to prevent