GATA-1 shRNA Plasmids (h)

shRNA gene silencing Human - TF‐1 GATA‐1

Experiment
shRNA gene silencing Human - TF‐1 GATA‐1
Product
GATA-1 shRNA Plasmids (h) from Santa Cruz Biotechnology
Manufacturer
Santa Cruz Biotechnology

Protocol tips

Protocol tips
TF‐1 cells (CRL‐2003; ATCC; Manassas, VA; RRID:CVCL_0559), an erythroleukemic cell line from human blood, were maintained at 37°C, 5% CO2 in RPMI 1640 medium supplemented with 10% (v/v) fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin, 1 mM sodium pyruvate, 4.5 g/L glucose and 2 ng/mL GM‐CSF (PHC2015). Prior to induction of differentiation, cells were kept in medium without GM‐CSF overnight, followed by the addition of EPO (1 U/mL; PHC2054) on the next day. The TF‐1 cell line is not listed in the database of commonly misidentified cell lines (International Cell Line Authentication Committee), and not authenticated by the authors. All culture reagents were from Life Technologies (Life Technologies, Carlsbad, CA, USA). The renewal of medium and EPO (1 U/mL) were done on alternative days, and the cell density was maintained at 4 × 105 cells/mL. Cultured TF‐1 cells were transfected with cDNA constructs with Trans IT X2® Dynamic Delivery System (Mirus BIO, Madison, WI, USA) for GATA‐1 shRNA and (Santa Cruz, Dallas, TX, USA). Transfection was performed 1 day after cells were plated. For every 5 × 106 cells in 1 mL medium, DNA: Trans IT X2® were mixed in the ratio of 1 : 2 (3 μg/6 μL) in antibiotic free medium and added to the cells within 20 min.

Publication protocol

TF‐1 cells (CRL‐2003; ATCC; Manassas, VA; RRID:CVCL_0559), an erythroleukemic cell line from human blood, were maintained at 37°C, 5% CO2 in RPMI 1640 medium supplemented with 10% (v/v) fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin, 1 mM sodium pyruvate, 4.5 g/L glucose and 2 ng/mL GM‐CSF (PHC2015). Prior to induction of differentiation, cells were kept in medium without GM‐CSF overnight, followed by the addition of EPO (1 U/mL; PHC2054) on the next day. The TF‐1 cell line is not listed in the database of commonly misidentified cell lines (International Cell Line Authentication Committee), and not authenticated by the authors. All culture reagents were from Life Technologies (Life Technologies, Carlsbad, CA, USA). The renewal of medium and EPO (1 U/mL) were done on alternative days, and the cell density was maintained at 4 × 105 cells/mL. Cultured TF‐1 cells were transfected with cDNA constructs with Trans IT X2® Dynamic Delivery System (Mirus BIO, Madison, WI, USA) for GATA‐1 shRNA and (Santa Cruz, Dallas, TX, USA). Transfection was performed 1 day after cells were plated. For every 5 × 106 cells in 1 mL medium, DNA: Trans IT X2® were mixed in the ratio of 1 : 2 (3 μg/6 μL) in antibiotic free medium and added to the cells within 20 min.

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

Reviews

GATA-1 shRNA Plasmids (h) from Santa Cruz Biotechnology has not yet been reviewed for this experiment

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

Discussion

Discussion

5 years ago

Author: Italy

Is a knockdown using shRNA permanent?

Is a knockdown using shRNA permanent and if not is there a known duration?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing shRNA gene silencing Human - TF‐1 GATA‐1 using GATA-1 shRNA Plasmids (h) from Santa Cruz Biotechnology.

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

Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for GATA-1 shRNA Plasmids (h) below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing shRNA gene silencing Human - TF‐1 GATA‐1 using GATA-1 shRNA Plasmids (h) from Santa Cruz Biotechnology. 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