Anti-Semaphorin 3A antibody (ab23393)

Western blot - Semaphorin 3A antibody (ab23393)

Lanes 1 - 2 : Anti-Semaphorin 3A antibody (ab23393) at 1/1000 dilution
Lanes 3 - 4 : Anti-Semaphorin 3A antibody (ab25999)

Lane 1 : Neonatal rat brain
Lane 2 : Human recombinant Sema3A/Fc chimera (95/125 kDa)
Lane 3 : Neonatal rat brain
Lane 4 : Human recombinant Sema3A/Fc chimera (95/125 kDa)


Predicted band size : 89 kDa


Membrane was incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody (ab23393)

Ab23393 staining human normal ileum. Staining is localized to the cytoplasm.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

Western blot - Semaphorin 3A antibody (ab23393)

Anti-Semaphorin 3A antibody (ab23393) at 1/1000 dilution + Lysate prepared from Xenopus laevis brain tissue section at 15 µg

Secondary
HRP-conjugated goat polyclonal to rabbit IgG at 1/20000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 89 kDa
Observed band size : 106 kDa (why is the actual band size different from the predicted?)
Additional bands at : 65 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 1 minute

This image is a courtesy of Anonymous Abreview

Immunohistochemistry (Frozen sections) - Anti-Semaphorin 3A antibody (ab23393)

ab23393 staining Semaphorin 3A in 16 µm thick sections of Apteronotus leptorhynchus brain, spinal cord by Immunohistochemistry (Frozen sections).

Tissue was fixed in 2% paraformaldehyde, permeabilized using 0.3% Triton X-100, then blocked with 3% sheep serum, 1% BSA, 1% teleostean gelatine in TBS for 1 hour at 24°C and then incubated with ab23393 at a 1/50 dilution for 18 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/200 dilution.

Image courtesy of Dr Ruxandra Sirbulescu by Abreview.