TCR beta Monoclonal Antibody (H57-597), FITC, eBioscience™

Flow cytometry Anti-bodies Mouse - TCRbeta

Experiment
Flow cytometry Anti-bodies Mouse - TCRbeta
Product
TCR beta Monoclonal Antibody (H57-597), FITC, eBioscience™ from eBioscience
Manufacturer
eBioscience

Protocol tips

Protocol tips
7AAD (Biolegend) or Zombie Aqua Fixable Viability Kit (Biolegend) was used to exclude dead cells. For intracellular transcription factor staining, surface-stained cells were fixed and permeabilized using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience), according to the manufacturer's protocol
Downstream tips
Cells were acquired on the BD FACSCANTO II and LSR (BD Biosciences) and analysis was carried out using FlowJo (Tree Star).

Publication protocol

Tumors tissues were harvested from mice that had been treated with mAb or otherwise and processed for flow cytometry analysis as previously described (20). For surface staining, tumor-filtrating leukocytes (TIL) were stained with eFluor780 anti-CD45.2 (104; eBioscience), eFluor450 or Brilliant Violet 605 anti-CD4 (RM4-5; eBioscience and Biolegend), PE-Cy7 or Brilliant Violet 421 anti-CD8a (53-6.7; eBioscience and Biolegend), FITC or PE anti-TCRβ (H57-597; eBioscience), PE-Cy7-anti-CD11b (M1/70; eBioscience), eFluor450-anti-Gr1 (RB6-8C5; eBioscience), FITC- or PE-anti-PD1 (J43; eBioscience and BD Pharmingen), APC-anti-PDL1 (10F.9G2; Biolegend), PE-anti-CD27 (LG.3A10; BD Pharmingen), PE-anti-CD86 (GL1; BD Pharmingen), APC-anti-CD80 (16-10A1; eBioscience), PE-anti-CD70 (FR70; BD Pharmingen), FITC-anti-CD40 (HM40-3; BD Pharmingen), Biotin-conjugated-anti-CD28 (37.51; Biolegend), PE-Cy7- or APC-conjugated streptavidin (eBioscience), Alexa Fluor 488-anti-CD25 (PC61.5; eBioscience), Brilliant Violet 605-anti-CD127 (A7R34; Biolegend), PE-Cy7-anti-CD278 (ICOS; 7E.17G9; eBioscience), APC-anti-CD223 (Lag3; C9B7W; Biolegend), PE-anti-CD366 (Tim3; RMT3-23; Biolegend), APC-anti-TIGIT (1G9; Biolegend), PE-Cy7 anti-CD39 (24DMS1; eBioscience), PE-anti-CD73 (TY/23; BD Pharmingen), APC-anti-CD44 (IM7; Biolegend), FITC-anti-CD62L (MEL-14; Biolegend), and respective isotype antibodies in the presence of anti-CD16/32 (2.4G2). 7AAD (Biolegend) or Zombie Aqua Fixable Viability Kit (Biolegend) was used to exclude dead cells. For intracellular transcription factor staining, surface-stained cells were fixed and permeabilized using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience), according to the manufacturer's protocol, and stained using eFluor450-anti-Foxp3 (FJK-16s, eBioscience), FITC-anti-Tbet (4B10; Biolegend), APC-anti-CTLA4 (UC10-4B9, eBioscience), Alexa Fluor 647-anti-Ki67 (B56), eFluor660-anti-Eomes (Dan11mag; eBioscience), and respective isotype antibodies. For intracellular staining of IFNγ/TNF or IL12p40, cells were stimulated in vitro with 50 ng/mL PMA (Sigma Aldrich) and 1 μg/mL ionomycin (Sigma Aldrich), or 100 ng/mL LPS, respectively in the presence of GolgiPlug (BD Biosciences) for 4 hours, and then surface stained as aforementioned. Surface-stained cells were then fixed and permeabilized using BD Cytofix/Cytoperm (BD Biosciences) according to the manufacturer's protocol, and stained with PE-anti-IL12p40 (C15.6; BD Pharmingen), PE-anti-IFNγ (XMG1.2; eBioscience), Alexa Fluor 647-anti-granzyme B (GB11; BD Pharmingen), and Brilliant Violet 605-anti-TNF (MP6-XT22; Biolegend), and respective isotype antibodies. Cells were acquired on the BD FACSCANTO II and LSR (BD Biosciences) and analysis was carried out using FlowJo (Tree Star).

Full paper   Login or join for free to view the full paper.

Reviews

TCR beta Monoclonal Antibody (H57-597), FITC, eBioscience™ from eBioscience has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Flow cytometry Anti-bodies Mouse - TCRbeta using TCR beta Monoclonal Antibody (H57-597), FITC, eBioscience™ from eBioscience.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from eBioscience for TCR beta Monoclonal Antibody (H57-597), FITC, eBioscience™ below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Flow cytometry Anti-bodies Mouse - TCRbeta using TCR beta Monoclonal Antibody (H57-597), FITC, eBioscience™ from eBioscience. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms