Anti-BAFF antibody [Buffy 2] (ab16081)

- Anti-BAFF antibody [Buffy 2] (ab16081)

Staining of paraffin section of synovial tissue from a patient with Rheumatoid Arthritis. Arrows indicate vessels (20x) staining positive for BAFF using ab16081.

Tissues are in paraffin sections. Sections are de-waxed and rehydrated, then cooked. Sections are stained in wet boxes, rehydrated and finally prepared as coverslip slides using Eukitt solution.

Immunocytochemistry/ Immunofluorescence - Anti-BAFF antibody [Buffy 2] (ab16081)

ICC/IF image of ab16081 stained HeLa cells. The cells were 4% PFA 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 (ab16081, 1µg/ml) overnight at +4ºC. 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) at a concentration of 1.43µM.

Flow Cytometry-BAFF antibody [Buffy 2](ab16081)

Overlay histogram showing THP1 cells stained with ab16081 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16081, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgGM (2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

Western blot

Anti-BAFF antibody [Buffy 2] (ab16081) at 1 µg/ml + BAFF protein (His tag) (ab78840) at 0.01 µg

Secondary
Peroxidase Conjugated AffiniPure Rabbit Anti-Rat IgG (H+L) (ab88508) at 1/10000 dilution
developed using the ECL technique

Performed under reducing conditions.

Exposure time : 4 minutes