Anti-Integrin beta 1 antibody [12G10]

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Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (AB30394)

ab30394 at 1/500 staining Integrin beta 1 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by Immunocytochemistry/ Immunofluorescence.

Cells were fixed in paraformaldehyde. The secondary used was an Alexa Fluor^® 488 Goat anti mouse (H + L), used at a 1/500 dilution. Actin filaments (Phalloidin-TRITC) and DNA (DAPI, blue) are also shown

This image is courtesy of an anonymous abreview.

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Flow Cytometry - Anti-Integrin beta 1 antibody [12G10] (AB30394)

Overlay histogram showing HAP1 wildtype (green line) and HAP1-ITGB1 knockout cells (red line) stained with ab30394.

Live HAP1 wildtype and HAP1-ITGB1 knockout cells were incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab30394, 1μg/0.5x10⁶ cells) for 30 min at 22°C. A mouse IgG1 isotype control antibody (ab170190) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-ITGB1 knockout - grey line). Unlabeled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

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Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (AB30394)

ab30394 staining Integrin beta 1 in wild-type HAP1 cells (top panel) and Integrin beta 1 knockout HAP1 cells (bottom panel).

The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab30394 at 10μg/ml concentration and ab202272 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labeled in blue with DAPI.

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Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (AB30394)

ab30394 staining Integrin beta 1 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab30394 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

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Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (AB30394)

ab30394 at 10 μg/ml staining Integrin beta 1 in HT1080 (Human fibrosarcoma cell line) cells by Immunocytochemistry/ Immunofluorescence.

Cells were fixed in paraformaldehyde. The secondary used was an Alexa Fluor^® 555 Donkey anti mouse (H + L), used at a 1/500 dilution.

This image is courtesy of an anonymous abreview.

• Flow Cyt

Lab

Flow Cytometry - Anti-Integrin beta 1 antibody [12G10] (AB30394)

Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab30394 (red line).

The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab30394, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight^® 488 (IgG; H+L) ab96879 at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

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CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (AB30394)

Immunocytochemistry-immunofluorescence using Anti-Integrin beta 1 antibody [12G10], ab30394. Publication image from Dorner, M. et al., 2018, Nat Commun, 29445209. Legend direct from paper.

3D PHH cultures are metabolically stable and exhibit functional tissue formation. a Comparison of cytochrome P450 gene (HNMT, CYP2A6, SLC22A1, CYP2A13, and GSTP1) mRNA expression between static 2D PHH cultures, 3D spheroid cultures, SACC PHH cultures, and 3D PHH cultures following 14 days of culture as well as freshly thawed PHH and HepG2 cells. b Longitudinal comparison of the mRNA expression profile of HNMT, CYP2A6, ABCB11, SLC22A17, CYP2A13, and GSTP1 as well as HBV receptors (NTCP, ASGPR) in HepG2 cells and freshly thawed PHH as well as static 2D and 3D PHH cultures. c Determination of CYP3A activity in six different hepatocyte donors 7 days following seeding in 3D PHH cultures as determined by CYP3A-Glo assay. d CYP450 activity in 3D PHH following 7, 14, or 21 days of culture time as determined by quantification of metabolites of Tacrine (CYP1A2), Diclofenac (CYP2C9), and Midazolam (CYP3A4) by quadrupole linear ion trap mass spectrometry. e Stable maintenance of the bile canaliculi marker DPPIV/CD26 in 3D PHH cultures as determined by Luminex. f Functionality of bile canaliculi as determined by CDFDA staining. g, h Ultrastructural analysis of 3D PHH by transmission electron microscopy after 20 days of culture. (Filled arrows : bile canaliculi, open arrows : hepatic microvilli, asterisk : tight junctions, L : lipid droplet). i Immunofluorescence microscopy of tight junction and hepatocyte markers (ZO-1, CD81, ITGB1, Connexin 32) in 3D PHH cultures following 26 days of culture. All data shown are mean ± SD of three to six independent experiments. Scale bars : white (200 µm), gray (2 nm), black (500 nm)