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AB263797

Human STAT3 knockout HeLa cell lysate

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STAT3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2.
4 Images
Western blot - Human STAT3 knockout HeLa cell lysate (AB263797)
  • WB

Lab

Western blot - Human STAT3 knockout HeLa cell lysate (AB263797)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

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 (20μg)
Lane 2 :****STAT3 knockout HeLa serum starved overnight, then treated with 50 ng/ml IFN alpha for 30 minutes, whole cell lysate (20μg)
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 (20μg)**
**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 (20μg)

Lanes 1-4 : Merged signal (red and green). Green - ab267373 observed at 88 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

Lanes 1-2 : 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 ab263797) was used. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. ab267373 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C 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 (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/stat3-phospho-y705-antibody-epr23968-52-ab267373'>ab267373</a>) 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

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 88 kDa

Observed band size: 88 kDa

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Western blot - Human STAT3 knockout HeLa cell lysate (AB263797)
  • WB

Lab

Western blot - Human STAT3 knockout HeLa cell lysate (AB263797)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : STAT3 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab109085 observed at 92 kDa. Red - loading control ab8245 observed at 37 kDa.

ab109085 Recombinant Anti-STAT3 antibody [EPR361] was shown to specifically react with STAT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255436 (knockout cell lysate ab263797) was used. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109085 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-STAT3 antibody [EPR361] (<a href='/en-us/products/primary-antibodies/stat3-antibody-epr361-ab109085'>ab109085</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

STAT3 knockout HeLa cell lysate at 20 µg

Predicted band size: 88 kDa

Observed band size: 92 kDa

false

Western blot - Human STAT3 knockout HeLa cell lysate (AB263797)
  • WB

Lab

Western blot - Human STAT3 knockout HeLa cell lysate (AB263797)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : STAT3 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab68153 observed at 92 kDa. Red - loading control ab8245 observed at 37 kDa.

ab68153 Recombinant Anti-STAT3 antibody [EPR787Y] was shown to specifically react with STAT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255436 (knockout cell lysate ab263797) was used. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab68153 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-STAT3 antibody [EPR787Y] (<a href='/en-us/products/primary-antibodies/stat3-antibody-epr787y-ab68153'>ab68153</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

STAT3 knockout HeLa cell lysate at 20 µg

Predicted band size: 88 kDa

Observed band size: 92 kDa

false

Sanger Sequencing - Human STAT3 knockout HeLa cell lysate (AB263797)
  • Sanger seq

Unknown

Sanger Sequencing - Human STAT3 knockout HeLa cell lysate (AB263797)

Homozygous : 1 bp deletion in exon2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2.

Disease

Adenocarcinoma

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Western blot - Human HK2 (Hexokinase II) knockout HEK-293 cell lysate (AB281353)

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Western blot - Human STAT1 knockout HeLa cell lysate (AB263837)

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Primary Antibodies

AB68153

Anti-STAT3 antibody [EPR787Y]

KO Validated|RabMAb|Recombinant|Lab Essentials|20ul selling size|Advanced Validation

Western blot - Anti-STAT3 antibody [EPR787Y] (AB68153)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
STAT3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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.
Biological function summary

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.

Pathways

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.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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