Immunofluorescence studies were carried out as previously described [24] (link). Briefly, cells grown overnight on 24-well cover glasses (Menzel, 12 mm) were treated with Cy2/Cy3-labeled MOA in FCS-containing medium and fixed in 4% paraformaldehyde (PFA) at 4 °C for 10 min. Cells were washed and incubated with NH4Cl at room temperature for 15 min before being permeabilized with PBS containing 0.02% saponin and 0.2% bovine serum albumin (BSA). Cells were stained with either anti-EEA1 (1:50, BD Transduction Laboratories), anti-Calnexin (1:100, Enzo Life Science), anti-CTR 433 (1:100, kind gift of Michel Bornens, Curie Institut), anti-Giantin (1:100, Abcam), anti-TGN46 (1:100, Sigma Aldrich), anti-Transferrin receptor (TfR, 1:100, Life Technologies), anti-Rab11 (1:100, Cell Signaling), anti-Lamp1 (1:200, BD PharMingen), anti-Rab7 (1:100, Santa Cruz Biotech) or anti-β1 integrin (1:100, R&D Systems) antibodies diluted in permeabilization buffer, followed by either donkey anti-mouse Cy3-labeled secondary antibody (1:100, Jackson ImmunoResearch), donkey anti-rabbit Cy3-labeled secondary antibody (1:100, Jackson ImmunoResearch) or donkey anti-goat Cy3-labeled secondary antibody (1:100, Jackson ImmunoResearch) diluted in permeabilization buffer. Nuclei were stained using DAPI (300 nM, Life Technologies).
Anti β1 integrin
Manufactured by R&D Systems
Anti-β1 integrin is a monoclonal antibody that specifically binds to the β1 subunit of integrin receptors. Integrins are cell surface receptors that mediate cell-cell and cell-extracellular matrix interactions and play a role in various cellular processes. The Anti-β1 integrin antibody can be used for the identification and study of β1 integrin expression and function.
Automatically generated - may contain errors
Lab products found in correlation
Celltiter glotm luminescent cell viability assay, by Promega (2 mentions)
Veritas microplate luminometer, by Promega (2 mentions)
Anti α1 integrin, by Merck Group (2 mentions)
Anti αv integrin, by Merck Group (2 mentions)
Anti eea1, by BD (1 mentions)
Anti calnexin, by Enzo Life Sciences (1 mentions)
Anti giantin, by Abcam (1 mentions)
Anti tgn46, by Merck Group (1 mentions)
Anti rab11, by Cell Signaling Technology (1 mentions)
Anti lamp1, by BD (1 mentions)
Anti rab7, by Santa Cruz Biotechnology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Anti integrin α2, by Merck Group (1 mentions)
Anti α3 integrin, by Merck Group (1 mentions)
Anti α5 integrin, by Merck Group (1 mentions)
Anti α6 integrin, by Merck Group (1 mentions)
4 protocols using anti β1 integrin
1
Immunofluorescence Assay for Organelle Markers
2
Integrin-Mediated Cell Adhesion Enhancement
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The involvement of integrins in the enhancement of cell adhesion by the ROCK inhibitor was evaluated by seeding MCECs (5 × 103 cells/well) in 96-well plates in the presence or absence of integrin-neutralizing antibodies (2 μg/mL): anti-α1 integrin (Merck Millipore, Billerica, MA), anti-α2 integrin (Merck Millipore, Billerica, MA), anti-α3 integrin (Merck Millipore, Billerica, MA), anti-α4 integrin (Merck Millipore, Billerica, MA), anti-α5 integrin (Merck Millipore, Billerica, MA), anti-α6 integrin (Merck Millipore, Billerica, MA), anti-αV integrin (Merck Millipore, Billerica, MA), anti-α6 integrin (Merck Millipore), and anti-β1 integrin (R&D systems Inc., Minneapolis, MN). The effect of inhibiting phosphorylation of MLC and RhoA activity on cell adhesion was also evaluated by seeding MCECs with blebbistatin (10 μM) and C3 (300 ng/ml), respectively. Three hours after seeding, the numbers of adherent cells were determined with the CellTiter-GloTM luminescent cell viability assay (Promega Corporation, Madison, WI) according to the manufacturer’s instructions. The number of adhered cells was determined using a VeritasTM microplate luminometer (Promega Corporation).
3
Quantification of VLA-4 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Expression of the Very Late Antigen-4 (VLA-4) receptor on the cell membrane of A2058 melanoma cells, K562 WT cells, and K562 VLA-4 expressing cells was determined using flow cytometry. In brief, cells were incubated with either anti-α4 or anti-β1 integrin (1:100 dilution; R&D Systems) in a solution containing 0.1% BSA for 2 hours on a rocker with ice. Cells were then washed three times with PBS and further incubated with the secondary antibody (1:1000 dilution; Invitrogen) for 2 hours on a rocker with ice. Finally, cells were washed three times and run through the flow cytometer. Analysis of the data was performed using Flow Jo.
4
Cell Adhesion on Extracellular Matrix
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The MCECs were seeded in 48-well or 96-well plates coated with or without PCM-DM. After 24 h, the mean cell areas were evaluated by Image J software (U.S. National Institutes of Health, Bethesda, MD) following actin staining. The numbers of adhered cells after 1 h of seeding were measured with a CellTiter-GloTM luminescent cell viability assay (Promega Corporation, Madison, WI) according to the manufacturer's instructions. The number of adhered MCECs at 1 h after seeding was then measured using a VeritasTM microplate luminometer (Promega Corporation). To elucidate the effect of the interaction between the integrins of the CECs and the PCM-DM on cell adhesion, the MCECs (1×104 cells/well) were seeded in 96-well plates coated with PCM-DM in the presence or absence of integrin-neutralizing antibodies (2 µg/mL), anti-α1 integrin (Merck Millipore, Billerica, MA), anti-αv integrin (Merck Millipore), and anti-β1 integrin (R&D systems Inc., Minneapolis, MN). After 3 h of seeding, adherent cells were measured with the CellTiter-GloTM luminescent cell viability assay.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!