All lanes : Anti-Bmi1 antibody [BMI1/2823] (ab269678)

Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : BMI1 knockout MCF7 cell lysate
Lane 3 : A431 cell lysate
Lane 4 : HEK-293 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 36 kDa
Observed band size: 36 kDa

Lanes 1- 4: Merged signal (red and green). Green - ab269678 observed at 36 kDa. Red - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) observed at 50 kDa.

 ab269678 was shown to react with Bmi1 in wild-type MCF7 cells in western blot. Loss of signal was observed when knockout cell line ab262319 (knockout cell lysate ab256851) was used. Wild-type MCF7 and BMI1 knockout MCF7 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab269678 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) overnight at 4°C at a 2 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for Bmi1 using ab269678 at 2 μg/ml in immunohistochemical analysis.

PFA-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Bmi1 using ab269678 at 2 μg/ml in ICC/IF. The secondary antibody is a Goat anti-mouse IgG-CF488 conjugate (Green). The membrane is stained with Phalloidin (Red).

Anti-Bmi1 antibody [BMI1/2823] (ab269678) at 2 µg/ml + NIH/3T3 (mouse embryo fibroblast cell line) cell lysate

Predicted band size: 36 kDa

Formalin-fixed, paraffin-embedded human prostate carcinoma tissue stained for Bmi1 using ab269678 at 2 μg/ml in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human colon carcinoma tissue stained for Bmi1 using ab269678 at 2 μg/ml in immunohistochemical analysis.

Analysis of Protein Array containing more than 19,000 full-length human proteins using ab269678.

Z- and S- Score: The Z-score represents the strength of a signal that a monoclonal antibody (MAb) (in combination with a fluorescently-tagged anti-IgG secondary antibody) produces when binding to a particular protein on the HuProt^TM array. Z-scores are described in units of standard deviations (SD’s) above the mean value of all signals generated on that array. If targets on HuProt^TM are arranged in descending order of the Z-score, the S-score is the difference (also in units of SD’s) between the Z-score. S-score therefore represents the relative target specificity of a MAb to its intended target. A MAb is considered to specific to its intended target, if the MAb has an S-score of at least 2.5. For example, if a MAb binds to protein X with a Z-score of 43 and to protein Y with a Z-score of 14, then the S-score for the binding of that MAb to protein X is equal to 29.