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AB280802

Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line

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PTGS2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

Cyclooxygenase, Cyclooxygenase 2b, Cyclooxygenase-2, EC 1.14.99.1, GRIPGHS, Glucocorticoid-regulated inflammatory Prostaglandin G/H synthase, Glucocorticoid-regulated inflammatory cyclooxygenase, Macrophage activation-associated marker protein P71/73, OTTHUMP00000033524, PES-2, PGG/HS, PGH synthase 2, PGH2_HUMAN, PGHS-2, PHS 2, PHS II, PTGS2, Prostaglandin G/H synthase, Prostaglandin G/H synthase 2, Prostaglandin G/H synthase 2 precursor, Prostaglandin G/H synthase and cyclooxygenase, Prostaglandin H2 synthase 2, Prostaglandin-endoperoxide synthase 2, TIS10, TIS10 protein, fj02a10, hCox 2, prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase), ptgs2a, unp1239, wu:fj02a10

4 Images
Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (AB280802)
  • WB

Lab

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (AB280802)

False colour image of Western blot : Anti-COX2 / Cyclooxygenase 2 antibody [EPR18377-106] - N-terminal staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab188183 was shown to bind specifically to COX2 / Cyclooxygenase 2. A band was observed at 74-76 kDa in wild-type A549 cell lysates with no signal observed at this size in PTGS2 knockout cell line ab280802 (knockout cell lysate ab283825). Band at 70 kDa in both wild-type and knockout samples is non-specific but exact protein is not determined. To generate this image, wild-type and PTGS2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR18377-106] - N-terminal (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr18377-106-n-terminal-ab188183'>ab188183</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PTGS2 knockout A549 cell lysate at 20 µg

Lane 3:

U-87 MG cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Observed band size: 74-76 kDa

false

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (AB280802)
  • WB

Lab

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (AB280802)

False colour image of Western blot : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab179800 was shown to bind specifically to COX2 / Cyclooxygenase 2. A band was observed at 75 kDa in wild-type A549 cell lysates with no signal observed at this size in PTGS2 knockout cell line ab280802 (knockout cell lysate ab283825). To generate this image wild-type and PTGS2 knockout A549 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr12012-ab179800'>ab179800</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PTGS2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (ab280802)

Lane 3:

U-87 MG cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Predicted band size: 69 kDa

false

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (AB280802)
  • WB

Lab

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (AB280802)

False colour image of Western blot : Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283574 was shown to bind specifically to COX2 / Cyclooxygenase 2. A band was observed at 74-76 kDa in wild-type A549 cell lysates with no signal observed at this size in PTGS2 knockout cell line ab280802 (knockout cell lysate ab283825). Band at 70 kDa in both wild-type and knockout samples is non-specific but exact protein is not determined. To generate this image, wild-type and PTGS2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-rm1026-ab283574'>ab283574</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PTGS2 knockout A549 cell lysate at 20 µg

Lane 3:

U-87 MG cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Observed band size: 74-76 kDa

false

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (AB280802)
  • WB

Lab

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (AB280802)

All lanes:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EP1978Y] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-ep1978y-ab62331'>ab62331</a>) at 1/500 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PTGS2 knockout A549 cell lysate at 20 µg

Lane 3:

U-87 MG cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Lane 5:

Wild-type Hela ab255929 cell lysate at 20 µg

Lane 6:

PTGS2 knockout HeLa ab255524 cell lysate at 20 µg

Secondary

Lanes 1 - 6:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 6:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 75 kDa

false

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Western blot

Disease

Carcinoma

Reactivity data

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

Product details

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.

What's included?

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Properties and storage information

Gene name
PTGS2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Handling procedures

Initial handling guidelines

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.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Cyclooxygenase 2 also known as COX2 is an enzyme involved in the conversion of arachidonic acid to prostaglandins which are lipid compounds with hormone-like effects. It has alternative names including prostaglandin-endoperoxide synthase 2. The molecular weight of COX2 is approximately 72 kDa. This enzyme is expressed in various tissues including the brain kidneys and areas of inflammation. COX2 expression increases during inflammatory responses and is induced by pro-inflammatory cytokines.
Biological function summary

COX2 plays a significant role in the inflammatory response and is part of the complex process of synthesizing prostaglandins. These compounds mediate inflammation and pain making COX2 an important target for understanding these processes. COX2 is not ubiquitously expressed but rather is induced in activated macrophages and other cells during inflammatory conditions. Its function is also important for normal physiological processes like ovulation and implantation.

Pathways

COX2 is essential in the prostaglandin biosynthesis pathway connecting it to the arachidonic acid metabolism pathway. Cyclooxygenase 2 works with phospholipase A2 which releases arachidonic acid from the phospholipid membrane. COX2 then converts this acid to prostaglandin H2 a precursor for other prostaglandins. COX1 the other isoform of cyclooxygenase is closely related to COX2 and while they have different expression patterns they share some functional similarities in these pathways.

COX2 is connected to inflammatory conditions like arthritis and cancer. Its expression often increases in various cancer types contributing to tumor growth and metastasis by promoting angiogenesis and suppressing immune responses. The enzyme is also linked to rheumatoid arthritis where its overexpression exacerbates inflammation. COX2 inhibitors like ketorolac tromethamine or naproxen structure mitigate symptoms by decreasing prostaglandin synthesis. These inhibitors also interact with COX1 but selective inhibition of COX2 targets inflammation more effectively with fewer gastric side effects associated with COX1 inhibition.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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