Depositors Institution: Laboratoire de Genetique et Physiologie du Developpement (LGPD) / CNRS
Positive Tested Species Reactivity: Drosophila
Depositors Notes: Fusion: 1986. Immunohistology: On whole mount Drosophila embryos, it decorates the extracellular matrix surrounding the pericardial cells and the cardial cells of the heart tube, from 10h embryogenesis onwards; it is synthesized and secreted by the pericardial cells; it also labels the ring gland and the oenocytes; no maternal expression; it is still present around the adult heart tube. Immunoblotting with EC11: three bands (200, 190, 165 kDa) differentially expressed during embryogenesis, the 165 kDa species being the only one to persist in the adult heart tube. Functional effects: In the absence of Pericardin, the heart epithelium appears disorganized during its migration in concert with the dorsal ectoderm. The alignment of the cardial and pericardial cells is interrupted by holes and both types of cells are found accumulated in clusters along the rows of cardioblasts. The protein is probably involved in the morphogenesis and in the maintenance of the heart epithelium during dorsal ectoderm closure. Immunohistochemistry: Pericardial cells, extracellular matrix surrounding the heart tube, ring gland, oenocytes.
Antigen Molecular Weight: Predicted: 165kDa; Apparent: 200, 190 and 165kDa
Immunogen: Nitrocellulose containing embryonic (10-14 hours Drosophila melanogaster, Oregon R embryos) proteins after western blot. The excised nitrocellulose band contained proteins migrating at the same level than myosin heavy chain.
Additional Information: Pericardin is a type IV collagen-like ECM protein contributing to the ECM layer that surrounds the myocardial–pericardial network. EC1 1 was isolated by chance in a screen for monoclonal antibodies recognizing the muscle myosin heavy chain [PMID 3117889]. There are three Uniprot entries for this protein, two (Q7YU48 and Q9U617) are supported by experimental evidence at transcript level; the third, Q9VTR6, is a predicted protein; all three proteins are quite different; Q9U617 was selected as Uniprot ID because it encodes the NFQSTYYTK peptide identified in [PMID 12070098] by immunoprecipitation and sequencing. In [PMID 12070098], EC11 was used for immunogold on EM.
Recommended Applications: Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
These hybridomas were created by your colleagues. Please acknowledge the hybridoma contributor and the Developmental Studies Hybridoma Bank (DSHB) in the Materials and Methods of your publications. Please email the citation to us.
For your Materials & Methods section:
EC11 anti-Pericardin was deposited to the DSHB by Gratecos, Danielle (DSHB Hybridoma Product EC11 anti-Pericardin)
Storage and Handling Recommendations
Although many cell products are maintained at 4°C for years without loss of activity, shelf-life at 4°C is highly variable.
To ensure retention of antibody activity, we recommend aliquotting the product into two parts:
1) a volume of antibody stored at 4°C to be used within two weeks.
2) the remaining product diluted with an equal volume of molecular grade glycerol and stored at -20°C.
While optimal Ig concentration for an application will vary, a good starting concentration for immunohistochemistry (IHC),
immunofluorescence(IF) and staining is 2-5 µg/ml.
For Western blots, the concentration is decreased by one order of magnitude (that is, 0.2-0.5 µg/ml).
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