Anti-Plexin A4 antibody

• WB

Unknown

Western blot - Anti-Plexin A4 antibody (AB39350)

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab39350 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Plexin A4 antibody (ab39350) at 1 µg/mL

Lane 1:

Brain (Mouse) Tissue Lysate at 20 µg

Lane 2:

Western blot - Mouse brain tissue lysate - total protein (0 days) (<a href='/en-us/products/tissue-lysates/mouse-brain-tissue-lysate-total-protein-0-days-ab7188'>ab7188</a>) at 20 µg

Lane 3:

Brain (Rat) Tissue Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/35000 dilution

Predicted band size: 212 kDa

Observed band size: 190 kDa,213 kDa,41 kDa,70 kDa

true

Exposure time: 8min

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Plexin A4 antibody (AB39350)

ICC/IF image of ab39350 stained human SH-SY5Y cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab39350, 1μg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

• WB

Project2712****

Western blot - Anti-Plexin A4 antibody (AB39350)

We are unsure as to the biological basis of the band at 190 kDa. However, it is successfully blocked by peptide (ab39349) suggesting this may be a possible isoform of Plexin A4.

All lanes:

Western blot - Anti-Plexin A4 antibody (ab39350) at 1/250 dilution

Lane 1:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 2:

Western blot - Mouse brain tissue lysate - total protein (0 days) (<a href='/en-us/products/tissue-lysates/mouse-brain-tissue-lysate-total-protein-0-days-ab7188'>ab7188</a>) at 10 µg

Lane 3:

Dorsal Root Ganglia (Mouse) Tissue Lysate at 10 µg

Lane 4:

Brain (Mouse) Tissue Lysate at 10 µg with Mouse Plexin A4 peptide (<a href='/en-us/products/unavailable/mouse-plexin-a4-peptide-ab39349'>ab39349</a>)

Lane 5:

Western blot - Mouse brain tissue lysate - total protein (0 days) (<a href='/en-us/products/tissue-lysates/mouse-brain-tissue-lysate-total-protein-0-days-ab7188'>ab7188</a>) at 10 µg with Mouse Plexin A4 peptide (<a href='/en-us/products/unavailable/mouse-plexin-a4-peptide-ab39349'>ab39349</a>)

Lane 6:

Dorsal Root Ganglia (Mouse) Tissue Lysate at 10 µg with Mouse Plexin A4 peptide (<a href='/en-us/products/unavailable/mouse-plexin-a4-peptide-ab39349'>ab39349</a>)

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 212 kDa

Observed band size: 190 kDa,213 kDa,63 kDa

false