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AB276121

Human RIPK1 knockout THP-1 cell line

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RIPK1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 77 bp deletion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
6 Images
Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)
  • WB

Lab

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)

All lanes:

Western blot - Anti-RIP antibody [EPR4689-100] (<a href='/en-us/products/primary-antibodies/rip-antibody-epr4689-100-ab178420'>ab178420</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Human RIPK1 knockout THP-1 cell line (ab276121)

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 75 kDa

Observed band size: 76 kDa

true

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)
  • WB

Lab

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)

All lanes:

Western blot - Anti-RIP antibody [EPR4689] (<a href='/en-us/products/primary-antibodies/rip-antibody-epr4689-ab125072'>ab125072</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Human RIPK1 knockout THP-1 cell line (ab276121)

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 75 kDa

Observed band size: 76 kDa

true

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)
  • WB

Lab

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)

Anti-RIPK1 antibody [EPR24883-85] (ab300617) 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, ab300617 was shown to bind specifically to RIPK1. A band was observed at 75 kDa in wild-type THP-1 cell lysates with no signal observed at this size in RIPK1 knockout cell line ab276121 (knockout cell lysate ab284221). To generate this image, wild-type and RIPK1 knockout THP-1 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-RIP antibody [EPR24883-85] (<a href='/en-us/products/primary-antibodies/rip-antibody-epr24883-85-ab300617'>ab300617</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Secondary

Lanes 1 - 4:

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

Lanes 1 - 4:

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

Predicted band size: 75 kDa

Observed band size: 75 kDa

false

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)
  • WB

Lab

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)

Western blot : Anti-RIPK1 antibody [EPR4689-100] (ab178420) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab178420 was shown to bind specifically to RIPK1. A band was observed at 75 kDa in wild-type THP-1 cell lysates with no signal observed at this size in RIPK1 knockout cell line. To generate this image, wild-type and RIPK1 knockout THP-1 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-RIP antibody [EPR4689-100] (<a href='/en-us/products/primary-antibodies/rip-antibody-epr4689-100-ab178420'>ab178420</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 75 kDa

Observed band size: 75 kDa

false

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)
  • WB

Lab

Western blot - Human RIPK1 knockout THP-1 cell line (AB276121)

Western blot : Anti-RIPK1 antibody [EPR4689] (ab125072) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab125072 was shown to bind specifically to RIPK1. A band was observed at 75 kDa in wild-type THP-1 cell lysates with no signal observed at this size in RIPK1 knockout cell line. To generate this image, wild-type and RIPK1 knockout THP-1 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-RIP antibody [EPR4689] (<a href='/en-us/products/primary-antibodies/rip-antibody-epr4689-ab125072'>ab125072</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 75 kDa

Observed band size: 75 kDa

false

Sanger Sequencing - Human RIPK1 knockout THP-1 cell line (AB276121)
  • Sanger seq

Supplier Data

Sanger Sequencing - Human RIPK1 knockout THP-1 cell line (AB276121)

77bp deletion in exon 3.

Key facts

Cell type

THP-1

Species or organism

Human

Tissue

Blood

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 77 bp deletion in exon 3

Disease

Acute Monocytic Leukemia

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Complementary Products

Primary Antibodies

AB300617

Anti-RIP antibody [EPR24883-85]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry - Anti-RIP antibody [EPR24883-85] (AB300617)

  • 4
  • 1
Cell Lines & Lysates

AB269489

Human TREM2 knockout THP-1 cell line

Advanced Validation

Western blot - Human TREM2 knockout THP-1 cell line (AB269489)

  • 0
  • 0
Cell Lines & Lysates

AB276092

Human TP53 (p53) knockout A549 cell line

Advanced Validation

Western blot - Human TP53 (p53) knockout A549 cell line (AB276092)

  • 0
  • 0
Cell Lines & Lysates

AB276116

Human CASP1 (Caspase-1) knockout THP-1 cell line

Advanced Validation

Western blot - Human CASP1 (Caspase-1) knockout THP-1 cell line (AB276116)

  • 0
  • 0
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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>" }, "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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.

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What's included?

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Properties and storage information

Gene name
RIPK1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
  • It is not recommended to allow the cell density to exceed 1x106 cells/mL.
Culture medium

RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol

Cryopreservation medium

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

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Supplementary information

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

RIP also known as Receptor-Interacting Protein or RIPK1 is a serine/threonine-protein kinase with a mass of approximately 74 kDa. It plays an important role in cell death and survival signaling pathways. RIP is expressed ubiquitously across various tissues indicating its importance in many cellular functions. The protein contains a kinase domain an intermediate domain for protein-protein interactions and a death domain which facilitates its involvement in apoptotic signaling processes.
Biological function summary

Receptor-Interacting Protein Kinase 1 (RIPK1) participates in regulating both necroptosis and apoptosis distinguishing itself as an important mediator in cell death mechanisms. As part of the necrosome complex which includes RIPK3 and MLKL RIPK1 functions in necroptosis—a programmed form of necrosis. This characteristic involvement shows its dual role in maintaining cell fate decisions making it an integral part of immune response and inflammation control.

Pathways

RIPK1 strongly associates with the TNF signaling pathway and NF-kB pathway. Its interaction with TNF receptor 1 (TNFR1) and consequent involvement with TRADD and TRAF2 mediates the signal transduction necessary for the activation of NF-kB leading to transcription of genes involved in survival and inflammation. This connection illustrates its capability to switch between promoting cell survival through NF-kB and facilitating cell death via necroptosis or apoptosis depending on cellular context and cues.

RIPK1 plays a significant role in conditions such as inflammatory diseases and neurodegenerative disorders. Its overactivation results in excessive cell death implicated in inflammatory conditions; necrostatin a necroptosis inhibitor targets RIPK1 to potentially mitigate this damage. Furthermore RIPK1's dysregulation links to Alzheimer's disease where it can interact with components like RIPK3 to exacerbate neurodegenerative processes. This relationship underlines the potential of targeting RIPK1 therapeutically to manage inflammation and neurodegeneration.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

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Product protocols

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Target data

See full target information RIPK1
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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'.

For licensing inquiries, please contact partnerships@abcam.com