Application
Flow Cytometry
Sample
Rat Cell (Spleen)
Permeabilization
Yes - eBiosciences Cat. No. 00-5523
Gating Strategy
Gating was performed on viable splenocytes (R1)
Specification
Spleen
Preparation
Cell harvesting/tissue preparation method: Rat splenocytes were isolated and erythrocytes were lyses by hypotonic lysis (Aqua Bidest + 2x PBS). Splenocytes were activated for 48 h by using Phytohaemagglutinin (PHA) 5 µg/ml. Splenocytes were washed in PBS (300 g, 10 min) and adjusted to 1x10^7 cells/ml in PBS + 2% FCS. 100µl cell suspension (1x10^6 cells) were stained with 10 µl (0.25 ug) ab104950 ROR gamma and analyzed by flow cytometry. Double staining was performed by using 0.5 µl (0.25 µg) CD4 FITC (eBioscience, 11-0040).
Sample buffer: Cells were washed, blocked and stained in PBS+2% FCS. For intracellular staining, a Transcription Factor Staining Buffer was used (eBiosciences Cat. No. 00-5523).
Sample buffer: Cells were washed, blocked and stained in PBS+2% FCS. For intracellular staining, a Transcription Factor Staining Buffer was used (eBiosciences Cat. No. 00-5523).
Fixation
eBiosciences Cat. No. 00-5523
Other product details
Incubation time
30 minute(s) · Temperature: 19°C · Diluent: PBS + 2% FCS
Secondary antibody
Secondary antibody
None used
Additional data
Additional Notes
Left panel of the figure shows gating on viable cells (R1). Histogram shows in black isotype control, blue unstained cells, red RORgammat antibody. Right panel shows in the upper figure isotype control, middle unstained cells, lower stained cells with RORgammat PE and CD4 FITC.
Abcam response
We would like to thank the customer for this valuable feedback in form of an Abreview.
So far, this antibody had not been tested with rat samples therefore we would like to encourage more customers to submit us their feedback on this sample type.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Abcam user community
Verified customer
投稿 Nov 04 2015