MSLN Antibody – Mouse Monoclonal

Monoclonal

IgG

MN-1

Mouse

Human

0.02 M potassium phosphate, 0.15 M sodium chloride, pH 7.2, 0.01% sodium azide, sterile filtered

Protein A Purified

1 mg/ml (lot specific)

Cross reactivity with homologues from other sources has not been tested.

Short term 4 degree C, long term aliquot and store at -20 degree C, avoid freeze thaw cycles.

67,940 Da

601051

ModBase 3D Structure for Q13421

Human

This antibody was produced in mice by repeated immunizations with a recombinant protein corresponding to the extracellular domain of human mesothelin.

This antibody has been tested for use in immunohistochemistry and western blotting. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 40 kD in size corresponding to mature mesothelin by western blotting in the appropriate cell lysate or extract. For immunohistochemistry, archival PEFF human tissues were deparaffinized followed by hydration. Antigen-retrieval is recommended. Block tissues with 1% BSA in PBS for 30 min at 23 degree C. Antibodies are diluted in 1% BSA and reacted with tissue for 60 min at room temperature.

MSLN/Mesothelin antibody was raised against Human

Recombinant protein

MSLN / Mesothelin antibody was raised against this antibody was produced in mice by repeated immunizations with a recombinant protein corresponding to the extracellular domain of human mesothelin.

Extracellular Domain

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