Anti-Integrin alpha 9 antibody [EPR9722]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 9 antibody [EPR9722] (AB140599)

Immunohistochemical analysis of paraffin embedded Human lung tissue labelling Integrin alpha 9 with ab140599 antibody at a dilution of 1/100.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 9 antibody [EPR9722] (AB140599)

Immunohistochemical analysis of paraffin embedded Human skin tissue labelling Integrin alpha 9 with ab140599 antibody at a dilution of 1/100.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Integrin alpha 9 antibody [EPR9722] (AB140599)

Western blot : Rabbit Monoclonal[EPR9722] to Integrin alpha 9 ab140599 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 100 kDa in Wild-type A549 cell lysates with no signal observed at this size in ITGA9 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 100pc LICOR in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Integrin alpha 9 antibody [EPR9722] (ab140599) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

ITGA9 knockout A549 at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 114 kDa,140 kDa

Observed band size: 100 kDa,95 kDa

false

• WB

Unknown

Western blot - Anti-Integrin alpha 9 antibody [EPR9722] (AB140599)

All lanes:

Western blot - Anti-Integrin alpha 9 antibody [EPR9722] (ab140599) at 1/1000 dilution

Lane 1:

NIH3T3 cell lysate at 10 µg

Lane 2:

A673 cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

A549 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 114 kDa

Observed band size: 140 kDa

false

• WB

CiteAb

Western blot - Anti-Integrin alpha 9 antibody [EPR9722] (AB140599)

Western Blotting using Anti-Integrin alpha 9 antibody [EPR9722], ab140599. Publication image from Sargent, M. A. et al., 2019, Nat Commun, 30622267. Legend direct from paper.

Thbs3 reduces surface integrin levels and destabilizes the sarcolemma. a Representative Western blots of sarcolemma protein extracts from hearts of tTA control, Thbs3 DTG, and Thbs4 DTG mice, assayed for integrin proteins and β-dystroglycan. Laminin2α2 and Cacna1c serve as loading controls. Quantitation is shown in Supplementary Figure 3a-k. The red box shows integrin proteins specifically reduced on the sarcolemma by Thbs3. b Western blots for input of microsomal protein extracts before immunoprecipitation for Thbs3 and β1D integrin and Cacna1c from NRVMs infected with Adβgal or AdThbs3 (left panel). The microsomal fraction was then immunoprecipitated (IP) for β1 integrin and blotted for the indicated proteins (right panel). Mouse IgG was used as a control for the IP. c Regimen of Evans blue dye (EBD) and isoproterenol (Iso, 300 mg/kg) injection to measure membrane permeability in the hearts of mice. d Representative histological heart sections from 3 tTA control and 3 Thbs3 DTG mice, imaged for WGA (green fluorescence) and leakage of EBD into the cardiomyocytes by red fluorescence subjected to the experimental regimen shown in c. Scale bars are 1 mm. e Regimen of Evans blue dye (EBD) injection after TAC surgery to measure membrane permeability in adult mice. Mice were sacrificed (Sac) 24 h after EBD injection, 1 week after TAC surgery. f, g Representative cardiac histological images of tTA control and Thbs3 DTG hearts 1 week after TAC surgery stained with WGA (green) and intracellular leakage of EBD (red) according to the experimental scheme shown in e, while g shows quantitation of EBD positive cells in the hearts of these mice. Scale bars are 100 µm. *P < 0.05 versus tTA control sham; two-tailed students T-test. Data are represented as percentage EBD positive cells (≥3000 cells from ≥8 animals). Error bars are +/− standard error of the mean and number of mice used is shown in the graph as individual data points

false