Flow Cytometry for Anti-Integrin beta 1 [IPI-TS2/16]
Addgene #252246
- Data Submitted By
- Zachary Anderson , Haiying Li , Travis Riedel , Haisun Zhu and Deborah Moshinsky
- Lab Name
- Institute for Protein Innovation (IPI)
- Submission Date
- March 25, 2026
- Publication Date
- March 27, 2026 (modified April 14, 2026)
- DOI
- https://doi.org/10.57733/addgene.e0b0mg
- Abstract
- Experimental results for Anti-Integrin beta 1 [IPI-TS2/16] (Addgene #252246) in a flow cytometry assay with ExpiCHO cells expressing human ITGB1.
- Citation
-
Flow Cytometry for Anti-Integrin beta 1 [IPI-TS2/16]. Anderson Z, Li H, Riedel T, Zhu H, Moshinsky D 2026. Addgene Report, https://doi.org/10.57733/addgene.e0b0mg
![Anti-Integrin beta 1 [IPI-TS2/16] (Addgene #252246) recognizes human ITGB1. Histogram from FACS analysis on ExpiCHO cells transfected with human ITGB1 (red), or B7H3 negative control (gray). Cells expressing human ITGB1 were labeled with Anti-Integrin beta 1 [IPI-TS2/16] and Alexa Fluor 647 F(ab’)2 goat anti-rabbit IgG Fc fragment (Jackson ImmunoResearch, 111-606-046). Labeled cells were analyzed with an Intellicyt iQue Screener Plus flow cytometer. Histograms were generated and normalized to mode using FlowJo™ v10.10.](https://datahub-media.addgene.org/report-images/e0b0mg/TS2-16_CellDisplay.png)
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 beta 1 [IPI-TS2/16]
Addgene #252246 -
Alexa Fluor 647 F(ab')2 goat anti-rabbit IgG Fc fragment
Jackson ImmunoResearch #111-606-046
Materials & Methods
Sample
- Target Species
- Human
- Cell / Tissue Type
- ExpiCHO cells
- Additional Information
- ExpiCHO cells were transiently transfected with human ITGB1 or B7H3 (negative control).
Buffer
- Name
- PBS
Primary Antibody
- Name
- Anti-Integrin beta 1 [IPI-TS2/16]
- Source
- Addgene
- Catalog Number
- 252246
- Host Species
- Rabbit
- Target Antigen
- ITGB1
- Concentration
- 15 µg/mL
- Time
- 30 min
- Temperature
- On ice
Secondary Antibody
- Name
- Alexa Fluor 647 F(ab')2 goat anti-rabbit IgG Fc fragment
- Source
- Jackson ImmunoResearch
- Catalog Number
- 111-606-046
- Concentration
- 2 µg/mL
- Time
- 30 min
- Temperature
- On ice
Additional Information
- Additional Information
EC50: A fourteen point titration of antibody concentrations, ranging from 660 nM (0.1 mg/mL) to 4.42 pM with a 1:2.5 dilution factor, against human and mouse ITGB1 showed reactivity towards human ITGB1 with an observed EC50 value of 1.88 nM.
Protocol: ExpiCHO cells were grown per manufacturer’s instruction. Plasmids expressing human ITGB1 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 primary antibodies.
- Primary antibody: 15 µg/mL in 1 mL Dulbecco's phosphate-buffered saline (DPBS), 0.5% BSA, with 0.9 mM Ca2+, 0.5 mM Mg2+, kept at 4 °C. 50 µL of antibody added to cells and incubated on ice for 30 min.
- Washing conditions: Cells pelleted and washed twice in 0.5% DPBS, 0.5% BSA, with 0.9 mM Ca2+, 0.5 mM Mg2+.
- Secondary antibody: 2 µg/mL Alexa Fluor 647 F(ab')2 goat anti-rabbit IgG Fc fragment (Jackson ImmunoResearch, 111-606-046) in 0.5% DPBS, 0.5% BSA, with 0.9 mM Ca2+, 0.5 mM Mg2+, 30 min, on ice.
- 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.
- Controls
- Control cells were transfected with plasmid expressing full-length human B7H3 together with the BFP plasmid.
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- Recognizes human ITGB1