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Western Immunoblotting Protocol (Primary Ab Incubation In BSA) |
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For Western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween-20 at 4ˇăC with gentle shaking, overnight.
A. Solutions and Reagents
NOTE: Prepare solutions with Milli-Q or equivalently purified water.
1. 1X Phosphate Buffered Saline (PBS) [moreˇ] |
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Immunohistochemistry Protocol (Paraffin) |
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IMPORTANT: See product data sheet for the appropriate wash buffer and antigen unmasking procedure.
• For Citrate/PBST, use steps 5a, 6a and C1.
• For Citrate/TBST, use steps 5b, 6a and C1.
• For EDTA/PBST, use steps 5a, 6b and C2. [moreˇ] |
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Immunofluorescence Protocol |
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IMPORTANT: Please refer to the APPLICATIONS section on the data sheet to determine IF THIS PRODUCT is validated and approved for the specific protocol you will be using.
A. Solutions and Reagents
NOTE: Prepare solutions with Milli-Q or equivalently purified water. [moreˇ] |
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Flow Cytometry Protocol for Intracellular Staining Using Conjugated Secondary Antibodies |
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A. Solutions and Reagents
1. 1X Phosphate Buffered Saline (PBS): Dissolve 8 g NaCl, 0.2 g KCl, 1.44 g Na2HPO4 and 0.24 g KH2PO4 in 800 mL distilled water (dH2O). Adjust the pH to 7.4 with HCl and the volume to 1 liter. Store at room temperature.
2. Formaldehyde (methanol free) [moreˇ] |
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Immunohistochemistry Troubleshooting |
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Weak or No Staining
Sources
Inadequate deparaffinization
Inactive primary antibodies
[moreˇ] |
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Troubleshooting Immunofluorescence Assays |
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Problem: Positive Control is negative or very weak
Possible Causes:
1. Microscope must be equipped with a filter system for FITC (maximum excitation wavelength 490 nm, mean emission wavelength 530 nm) and 400X magnification.
[moreˇ] |
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Western Blotting Troubleshooting |
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Problem: Poor transfer quality: The poor transfer of proteins to the membrane can be visualized by staining the gel after transfer. [moreˇ] |
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Antibody Products FAQ |
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The high quality of our products is the most important consideration at EnoGene. Please contact EnoGene technical support at service@enogene.com for the latest application update or for help with any questions you may have. [moreˇ] |
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Custom Polyclonal Antibody Production FAQ |
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Customized protocols can always be developed, however, we have documented a greater than 95% success rate using the protocol listed below
[moreˇ] |
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