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Western blot
ImmunohistochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Immunohistochemistry [1]
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Validation data
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- Product number
- MAB3318 - Provider product page
- Provider
- EMD Millipore
- Proper citation
- Millipore Cat#MAB3318, RRID:AB_94813
- Product name
- Anti-TIMP-3 Antibody, a.a. 170-188, clone 136-13H4
- Antibody type
- Monoclonal
- Antigen
- Oligopeptide of SWYRGWAPPDKSIINATDP (residue 170-188) on human tissue inhibitor of metalloproteinases-3.
- Reactivity
- Human, Rabbit
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 136-13H4
- Vial size
- 100 µg
- Storage
- Maintain for 1 year at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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Enhanced validation
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- 59f2f45fd1842
- Enhanced method
- Independent antibody validation
- Main image
- Experimental details
- Formalin-fixed human post-mortem brain tissue from BA9. Tissue sample is from patient with CADASIL, a vascular condition that has been demonstrated to lead to accumulated TIMP3 and Notch3 around vessels. Tissue was dewaxed in xylene and re-hydrated in graded alcohols. Sections underwent heat-mediated antigen retrieval in DIVA decloaker buffer (DV2004, Biocare Medical) using a Decloaking Chamber (Biocare Medical) for 20 minutes at 11 0oC. Tissue was then permeabilised in tris-buffered saline (Sigma-Aldrich) containing 0.2% Tween 20 for 10 minutes at room temperature. Non-specific signal was reduced by blocking in 10% goat serum and 1% BSA in TBS (Sigma-Aldrich) for two hours at room temperature. Tissue was incubated overnight with primary antibodies anti-Notch3 1E4 (0.005 mg/ml, MABC594, EMD Millipore) or anti-TIMP3 (0.01 mg/ml, MAB3318, EMD Millipore) overnight in diluent solution (1% BSA in TBS) at 4oC. Tissue was washed three times for five minutes with TBS (Sigma-Aldrich) and appropriate Alexa Fluor (R) IgG secondary antibodies (Invitrogen) were applied for 1 h at 37oC and protected from light. Tissue was mounted using Vectashield without DAPI (H-1000, Vector laboratories) and 1.5 coverslips (Menzel-Gläser). Slides were imaged using fluorescent microscopy (Nikon).
