Anti-beta 2 Adrenergic Receptor antibody (ab36956)

Western blot - beta 2 Adrenergic Receptor antibody (ab36956)

All lanes : Anti-beta 2 Adrenergic Receptor antibody (ab36956) at 1 µg/ml

Lane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 4 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466) with beta 2 Adrenergic Receptor peptide (ab38102) at 1 µg/ml
Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179) with beta 2 Adrenergic Receptor peptide (ab38102) at 1 µg/ml
Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate with beta 2 Adrenergic Receptor peptide (ab38102) at 1 µg/ml

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 47 kDa
Observed band size : 55 kDa (why is the actual band size different from the predicted?)


Exposure time : 30 seconds

Flow Cytometry - beta 2 Adrenergic Receptor antibody (ab36956)

ab36956 staining the beta 2 adrenergic receptor in the Human AML cell line Monomac6 by Flow Cytometry. Cells were cultured in RPMI medium supplemented with antibiotics, L-glutamin, minimal essential amino acids and sodium pyruvate in PBS + 1% FCS and 0.01% sodium azide. The sample was incubated with the primary antibody (1/50 in PBS + 1% FCS and 0.01% sodium azide) for 30 minutes at 4ºC. A phycoerythrin-conjugated Donkey anti-rabbit IgG (1/100) was used as the secondary antibody.
Gating Strategy: Live Cells.

This image is courtesy of an Abreview submitted by Alexander Kalinkovich

Immunocytochemistry/ Immunofluorescence - beta 2 Adrenergic Receptor antibody (ab36956)

ICC/IF image of ab36956 stained PC12 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 (ab36956, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in 100% methanol fixed (5 min) PC12 cells at 5µg/ml.