Anti-Semaphorin 7a antibody
| Name | Anti-Semaphorin 7a antibody |
|---|---|
| Supplier | Abcam |
| Catalog | ab23578 |
| Prices | $400.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | WB ICC/IF ICC/IF IHC-F IHC-F IHC-P IP |
| Species Reactivities | Mouse, Rat, Bovine |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Mouse Semaphorin 7a |
| Blocking Peptide | Mouse Semaphorin 7a peptide |
| Description | Rabbit Polyclonal |
| Gene | Sema7a |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
ICC/IF image of ab23578 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23578, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
ab23578 detecting Semaphorin 7a in developing mouse neurons, differentiated from neural precursor cells cultured from embryonic day 13 neocortex; neurons were cultured on poly-D-lysine in the presence of Neurobasal Medium containing B27 supplement. Immunocytochemistry: all steps were performed in PBS. Cultures were fixed in 4% PFA for 15min, permeabilised with 0.1% Triton X-100 for 10min and blocked with 5% BSA, 0.1% Triton X-100 for 45min. ab23578 was incubated at 5µg/ml (12h in 5% BSA, 0.1% Triton X-100) at 4°C. Goat anti-rabbit AlexaFluor 488 was used as secondary antibody at 1/400 for 1hr at RT (in 5% BSA, 0.1% Triton X-100). Treated cultures were mounted on glass coverslips with Mowiol. ab23578 immunostaining (green colour) was neuronal and for the most part at the membrane with some cytosolic staining observed. No nuclear staining was observed, confirmed with a nuclear counterstain (To-pro-3; data not shown).
All lanes : Anti-Semaphorin 7a antibody (ab23578) at 1 µg/mlLane 1 : Brain (Mouse) Tissue Lysate (ab27253)Lane 2 : Brain (Mouse) Tissue Lysate - normal tissue, 0 days old (ab7188)Lane 3 : Brain (Mouse) Tissue Lysate (ab27253) with Mouse Semaphorin 7a peptide (ab30844) at 1 µg/mlLane 4 : Brain (Mouse) Tissue Lysate - normal tissue, 0 days old (ab7188) with Mouse Semaphorin 7a peptide (ab30844) at 1 µg/mlLysates/proteins at 20 µg per lane.SecondaryRabbit IgG secondary antibody (ab28446) at 1/10000 dilutionPerformed under reducing conditions.
![Semaphorin 7a was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to Semaphorin 7a and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab23578.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 75kDa: Semaphorin 7a.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/28190_Semaphorin-7a-Primary-antibodies-ab23578-5.jpg)
Semaphorin 7a was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to Semaphorin 7a and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab23578.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 75kDa: Semaphorin 7a.
Product References
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Caspase-9 activation revealed by semaphorin 7A cleavage is independent of - Caspase-9 activation revealed by semaphorin 7A cleavage is independent of
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