Recombinant Mouse Anti-Integrin alpha 4 Antibody (ACT-1)

SDS-PAGE analysis of PABX-122-FcS in reduced (Lane 1) and non-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result, different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

The purity of PABX-122-FcS was greater than 95% as determined by SDS-PAGE.

The purity of PABX-122-FcS was greater than 95% as determined by SEC-HPLC.

Western blot analysis of PABX-122-FcS was performed by loadingIntegrin alpha 4&beta Heterodimer Protein. Integrin alpha 4 migrates as 60-70 kDa, 80-85 kDa and 135-150 kDa

PABX-122-FcS incubation concentration: 2 ng/μL.
The secondary antibody: HRP-anti-mouse IgG
Lane 1: Reducing Antigen (1 μg)
Lane 2: Reducing Antigen (2 μg)

ELISA analysis of PABX-122-FcS was performed by coating with Integrin alpha 4&beta Heterodimer Protein.

The secondary antibody: HRP-anti-mouse IgG

RPMI8866 cells (0.5 x10⁶) were incubated with 1 μg APC labeled ACT-1 (α4β7 specific antibody). Antibody binding was measured by flow cytometry and is presented as a histogram. Over 99% of the RPMI8866 cells expressed α4β7 integrin. Unstained cells (grey shading) and isotype control antibody (solid line) were used as controls.

Peachman, K. K., Karasavvas, N., Chenine, A. L., McLinden, R., Rerks-Ngarm, S., Jaranit, K., ... & Michael, N. L. (2015). Identification of new regions in HIV-1 gp120 variable 2 and 3 loops that bind to α4β7 integrin receptor. PLoS One, 10(12), e0143895.

RMPI8866 cells were incubated with media only (solid circles) or with ACT-1 (0.5 μg/well; open circles, left panel) and then added to MAdCAM-1 (natural ligand of α4β7 integrin; 0.2 μg/well) coated plates. The plates were washed and 100 μl of media and 10 μl of AlamarBlue® dye were added to each well. Fluorescence was measured immediately after the addition of the dye (time 0) and then at 2 hours intervals for 8 hours. The percent inhibition at the 8 hour time point is shown.

Peachman, K. K., Karasavvas, N., Chenine, A. L., McLinden, R., Rerks-Ngarm, S., Jaranit, K., ... & Michael, N. L. (2015). Identification of new regions in HIV-1 gp120 variable 2 and 3 loops that bind to α4β7 integrin receptor. PLoS One, 10(12), e0143895.

RMPI8866 cells were incubated with media (solid circles), ACT-1 (open circles) and then added to (B) linear (left panel), cyclic V2 peptides (TH023, MN, and acute C, second, third and right panels, respectively), or (C) cyclic V3 peptides (TH023, left panel, MN, middle panel, and acute C, right panel) coated plates respectively. The plates were washed and 100 μl of media and 10 μl of AlamarBlue® dye was added to each well. Fluorescence was measured immediately after the addition of the dye (time 0) and then at 2 hour intervals for 8 hours. The percent inhibition at the 8 hour time point is shown in each panel.

Peachman, K. K., Karasavvas, N., Chenine, A. L., McLinden, R., Rerks-Ngarm, S., Jaranit, K., ... & Michael, N. L. (2015). Identification of new regions in HIV-1 gp120 variable 2 and 3 loops that bind to α4β7 integrin receptor. PLoS One, 10(12), e0143895.