Western blot - BMP4 antibody (ab39973)

All lanes : Anti-BMP4 antibody (ab39973) at 1 µg/ml

Lane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)
Lane 2 : Brain (Mouse) Tissue Lysate
Lane 3 : Brain (Rat) Tissue Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution

Predicted band size : 47 kDa
Observed band size : 47 kDa
Additional bands at : 42 kDa. We are unsure as to the identity of these extra bands.

Western blot - BMP4 antibody (ab39973)

All lanes : Anti-BMP4 antibody (ab39973) at 1 µg/ml

Lane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)
Lane 2 : Brain (Mouse) Mouse Tissue Lysate
Lane 3 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)
Lane 4 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)
Lane 5 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330)
Lane 6 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466) with BMP4 peptide (ab40140) at 1 µg/ml
Lane 7 : Brain (Mouse) Mouse Tissue Lysate with BMP4 peptide (ab40140) at 1 µg/ml
Lane 8 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889) with BMP4 peptide (ab40140) at 1 µg/ml
Lane 9 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431) with BMP4 peptide (ab40140) at 1 µg/ml
Lane 10 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330) with BMP4 peptide (ab40140) at 1 µg/ml

Lysates/proteins at 10 µg per lane.

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

Performed under reducing conditions.

Predicted band size : 47 kDa
Observed band size : 47 kDa
Additional bands at : 100 kDa. We are unsure as to the identity of these extra bands.

ab39973 detects a 47kDa band that we believe corresponds to the pre-cleaved form of BMP4 with the immunizing peptide raised against a region located in the propeptide. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

Western blot - BMP4 antibody (ab39973)

All lanes : Anti-BMP4 antibody (ab39973) at 1/1000 dilution

Lane 1 : mouse brain
Lane 2 : mouse brain
Lane 3 : mouse brain
Lane 4 : mouse brain

Lysates/proteins at 20 µg per lane.

Secondary
Dako Cytomation P0260 Conjugation: HRP at 1/3000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 47 kDa


Exposure time : 1 minute

Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom

Immunocytochemistry/ Immunofluorescence - BMP4 antibody (ab39973)

ICC/IF image of ab39973 staining primary hippocampal neurons. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 5% normal donkey serum / 0.3M glycine in 0.1% PBS-Tritonx100 for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab39973, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488donkey anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom