STAT3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2.
HeLa
Human
Cervix
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2
1110034C02Rik, ADMIO, APRF, AW109958, Acute-phase response factor, DNA binding protein APRF, FLJ20882, HIES, MGC16063, STAT3_HUMAN, Signal transducer and activator of transcription 3, Signal transducer and activator of transcription 3 (acute phase response factor)
STAT3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2.
HeLa
Human
Cervix
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2
Adenocarcinoma
STAT3
Knockout
CRISPR technology
Sanger Sequencing, Western blot
Homozygous
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
This supplementary information is collated from multiple sources and compiled automatically.
Signal Transducer and Activator of Transcription 3 (STAT3) also called p-stat3 or phospho-STAT3 is a critical transcription factor involved in various cellular processes. The molecular weight of STAT3 is approximately 92 kDa. This protein is expressed broadly across many tissues and cell types. Mechanically upon cytokine or growth factor stimulation STAT3 undergoes phosphorylation resulting in dimerization. This phosphorylated form often referred to as phospho-Stat3 or p-STAT3 translocates to the nucleus where it binds specific DNA sequences to modulate gene transcription.
STAT3 is an important player in regulating cell growth and apoptosis. It functions not only as a transcription factor but also as part of a larger protein complex that operates within the cell nucleus to influence gene expression. This multifaceted role enables STAT3 to control the expression of genes related to cell proliferation survival and differentiation impacting various biological processes. These functions highlight its importance in tissue homeostasis and response to extracellular signals.
STAT3 is actively involved in the JAK-STAT signaling pathway a principal route for many cytokines and growth factors and the MAPK pathway. STAT3 interacts with proteins such as JAK kinases which phosphorylate Stat3 and initiate the STAT3 signaling cascade. Additionally it closely associates with the protein Raf1 in the MAPK pathway linking external signals to transcriptional responses. These pathways play a significant role in immune response inflammation and growth signaling.
Researchers have implicated STAT3 in cancer and autoimmune diseases. Persistent activation of STAT3 is often observed in various cancers including breast and lung cancer promoting oncogenesis through altered gene expression. Additionally abnormal STAT3 signaling is associated with autoimmune conditions such as rheumatoid arthritis. The interplay between STAT3 and proteins like IL-6 in these diseases highlights its potential as a therapeutic target for intervention.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
Lanes 1-4: Merged signal (red and green). Green - Anti-STAT3 (phospho Y705) antibody [EPR23968-52] ab267373 observed at 88 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.
Lanes 1-2: Anti-STAT3 (phospho Y705) antibody [EPR23968-52] ab267373 Anti-STAT3 (phospho Y705) antibody [EPR23968-52] was shown to specifically react with STAT3 in wild-type serum starved and then IFN alpha treated HeLa cells. Loss of signal was observed when serum starved and then IFN alpha treated knockout cell line ab255436 (knockout cell lysate Human STAT3 knockout HeLa cell lysate ab263797) was used. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Anti-STAT3 (phospho Y705) antibody [EPR23968-52] ab267373 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at 4° overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
Lysates loaded onto lanes 3-4 were made freshly and used in WB immediately to minimize protein degradation.
All lanes: Western blot - Anti-STAT3 (phospho Y705) antibody [EPR23968-52] (Anti-STAT3 (phospho Y705) antibody [EPR23968-52] ab267373) at 1/1000 dilution
Lane 1: Wild-type HeLa (human cervix adenocarcinoma epithelial cell) serum starved overnight, then treated with 50 ng/ml IFN alpha for 30 minutes, whole cell lysate at 20 µg
Lane 2: STAT3 knockout HeLa serum starved overnight, then treated with 50 ng/ml IFN alpha for 30 minutes, whole cell lysate at 20 µg
Lane 3: Jurkat (human t cell leukemia cell line from peripheral blood) treated with 50 ng/ml IFN alpha for 30 minutes, whole cell lysate at 20 µg
Lane 4: HepG2 (human hepatocellar carcinoma epithelial cell) serum starved overnight, then treated with 100 ng/ml IL-6 for 30 minutes, whole cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution
Predicted band size: 88 kDa
Observed band size: 88 kDa
Homozygous: 1 bp deletion in exon 2.
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com