Anti-Integrin alpha 5 beta 1 [IPI-ITGA5/ITGB1.4] in Flow Cytometry (Human)
Addgene #213652
- Data Submitted By
- IPI Staff
- Lab Name
- Institute for Protein Innovation (IPI)
- Submission Date
- March 20, 2024
- Publication Date
- March 25, 2024 (modified March 27, 2024)
- Abstract
- Experimental results for Anti-Integrin alpha 5 beta 1 [IPI-ITGA5/ITGB1.4] (Addgene Antibody 213652) in a flow cytometry assay using ExpiCHO-S cells transiently transfected with human RGD-binding integrin α5β1.
Image attribution: Institute for Protein Innovation (IPI)
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-Integrin alpha 5 beta 1 [IPI-ITGA5/ITGB1.4]
Addgene #213652 -
Alexa Fluor 647 F(ab')2 fragment goat anti-rabbit IgG, Fc fragment-specific
Jackson ImmunoResearch #111-606-046
Materials & Methods
Sample
- Species
- Human
- Cell / Tissue Type
- ExpiCHO-S cells
- Additional Information
- ExpiCHO-S cells were transiently transfected with BFP and either human B7H3 (Control), human RGD-binding integrin α5β1, or mouse RGD-binding integrin α5β1.
Primary Antibody
- Name
- Anti-Integrin alpha 5 beta 1 [IPI-ITGA5/ITGB1.4]
- Source
- Addgene
- Catalog Number
- 213652
- Species
- Rabbit
- Gene / Target
- ITGA5, ITGB1
- Concentration
- 10 µg/mL
- Time
- 30 min
- Temperature
- 4 ˚C
Secondary Antibody
- Name
- Alexa Fluor 647 F(ab')2 fragment goat anti-rabbit IgG, Fc fragment-specific
- Source
- Jackson ImmunoResearch
- Catalog Number
- 111-606-046
- Concentration
- 1:150
- Time
- 30 min
- Temperature
- 4 ˚C
Additional Information
- Additional Information
ExpiCHO-S cells were grown in ExpiCHO expression medium (ThermoFisher Scientific) per manufacturer’s instruction. Integrin α and β subunits were cloned in separate expression plasmids and were transiently transfected into cells together with a plasmid expressing blue fluorescent protein (BFP). Control cells were transfected with plasmid expressing full-length human B7H3 together with the BFP plasmid. Two days after transfection, cells at a density of 3 x 106 cells/mL were labeled with the rabbit chimeric primary antibody.
Washing conditions: The cells were washed twice in 0.5% DPBS, 0.5% BSA, with 0.9 mM Ca2+, 0.5 mM Mg2+.
Detection: Labeled cells were analyzed with an Intellicyt iQue Screener Plus flow cytometer. Cells were first gated for positive BFP expression and then gated for the Alexa 647-conjugated secondary antibody.
Results
- Result
- Pass (The antibody worked under these conditions)