Anti-His Tag Antibody Picoband® (monoclonal, 19b10)

Western blot analysis of His Tag using anti-His Tag antibody (M30975-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: recombinant mouse His Tag protein 10ng,
Lane 1: recombinant mouse His Tag protein 5ng,
Lane 1: recombinant mouse His Tag protein 2.5ng,
Lane 1: recombinant mouse His Tag protein 1.25ng.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-His Tag antigen affinity purified monoclonal antibody (Catalog # M30975-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system.

Purification and identification of rLP78. (A) SDS-PAGE analysis of the purification of rLP78 by Ni + column. Lane M, protein molecular weight marker; Lane 1, the supernatant of the whole bacterial lysate; Lane 2, elution with 50 mM of imidazole; Lane 3, elution with 500 mM of imidazole. (B) Western blotting analysis of the purified rLP78 using anti-His-tag monoclonal antibody. (C) Western blotting analysis of the purified rLP78 using mouse anti- M. synoviae polyclonal antibodies.
Index in PubMed under a CC BY license. PMID: 38264482

Activity of rLP78 to adhere to membrane proteins of DF-1 cells. Microtiter ELISA plates were coated with the extracted membrane protein, cytosolic protein or BSA. (A) Different concentrations of rLP78 were added to individual wells. Bound proteins were detected using a mouse anti-His-tag monoclonal antibody. (B) Adherence of rLP78 (100 μg/mL) to membrane proteins of DF-1 cells was inhibited by mouse anti-LP78 serum and further detected using goat anti-mouse IgG. Bars represent the mean ± standard deviation of the OD 450nm values of samples in triplicate. The differences were compared between each group. The same letter indicates no obvious difference ( p > 0.05); different letters indicate a significant difference ( p < 0.05).
Index in PubMed under a CC BY license. PMID: 38264482

Binding ability of rLP78 to human fibronectin (hFn) and human plasminogen (hPlg). The binding ability of rLP78 to hFn (A) or hPlg (B) was identified by ELISA. The plates were coated with 5 μg/mL of hFn or hPlg. Different concentrations of rLP78 or BSA were added to individual wells. Bound proteins were detected using a mouse anti-His-tag monoclonal antibody. Bars represent the mean ± standard deviation of the OD values of samples in triplicate. The differences were compared between each group. The same letter indicates no obvious difference ( p > 0.05); different letters indicate a significant difference ( p < 0.01). Binding ability of rLP78 to hFn (C) or hPlg (D) was confirmed by western blotting. Bound hFn was determined by rabbit anti-hFn antibody or rabbit anti-plasminogen antibody. BSA was chosen as negative control.
Index in PubMed under a CC BY license. PMID: 38264482