All lanes : Anti-Macrophage inflammatory protein 1 alpha/CCL3+CCL3L1 antibody [EP493Y] (ab46786) at 1/2000 dilution

Lane 1 : Human Macrophage Inflammatory Protein 1 alpha / CCL3 active protein
Lane 2 : Human Macrophage Inflammatory protein 1 beta / CCL4 active protein
Lane 3 : Human Macrophage Inflammatory active protein 4/CCL18
Lane 4 : Human CCL3L1 recombinant protein

Lysates/proteins at 0.02 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 10 kDa
Observed band size: 10, 40 kDa why is the actual band size different from the predicted?


Exposure time: 10 seconds

Human CCL3 active protein contains aa27-92.

Human CCL4 active protein contains aa24-92.

Human CCL18 active protein contains aa21-89.

Human CCL3L1 recombinant protein contain 2*(aa28-93) sequence with His-tag and GST-tag.

Blocking and diluting buffer: 5% NFDM/TBST.

All lanes : Anti-Macrophage inflammatory protein 1 alpha/CCL3+CCL3L1 antibody [EP493Y] (ab46786) at 1/1000 dilution

Lane 1 : Untreated THP-1 (Human monocytic leukemia cell line) lysate
Lane 2 : THP-1 lysate treated with 100 nM Phorbol-12-myristate-13-acetate (PMA) overnight, then treated with Lipopolysaccharides (LPS) at 100 ng/ml for 4 hour and then 1 µg/ml Brefeldin A was added for the last 3 hours.

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 10 kDa
Observed band size: 10 kDa


Exposure time: 3 minutes

Blocking and diluting buffer: 5% NFDM/TBST.

Macrophage inflammatory protein 1 alpha is induced in macrophages following exposure to bacterial lipopolysaccharide (LPS), PMID: 9848081.