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AB265052

Human MAPK1 (ERK2) knockout HeLa cell line

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(1 Publication)

MAPK1 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and Insertion of the selection cassette in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
7 Images
Immunocytochemistry/ Immunofluorescence - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)

ab32081 staining ERK2 in wild-type HeLa cells (top panel) and ERK2 knockout HeLa cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab32081 at 0.2µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

Sandwich ELISA - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)
  • sELISA

Supplier Data

Sandwich ELISA - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)

Sandwich ELISA of ab314784 with the capture antibody dilution at 2 µg/mL and detector antibody dilution at 0.5 µg/mL. Interpolated concentrations of native ERK2 in human control wild type HeLa cell and MAPK1 (ERK2) knockout HeLa cell based on 500 µg/mL extract loads. The concentrations of ERK2 were measured in duplicate and interpolated from the ERK2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ERK2 concentration was determined to be 8032.4 pg/mL in wild type HeLa extract (Human wild-type HeLa cell line ab255448) and undetectable in MAPK1 (ERK2) knockout HeLa extract (Human MAPK1 (ERK2) knockout HeLa cell line ab265052).

Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)
  • WB

Lab

Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)

Lanes 1-2 : Merged signal (red and green). Green - ab184699 observed at 44 kDa. Red - loading control ab8245 observed at 37 kDa.

ab184699 Anti-ERK1 + ERK2 antibody [EPR17526] was shown to specifically react with ERK2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265052 (knockout cell lysate ab257525) was used. Wild-type and ERK2 knockout samples were subjected to SDS-PAGE. ab184699 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 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-ERK1 + ERK2 antibody [EPR17526] (<a href='/en-us/products/primary-antibodies/erk1-erk2-antibody-epr17526-ab184699'>ab184699</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MAPK1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (ab265052)

Predicted band size: 41 kDa

Observed band size: 44 kDa

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Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)
  • WB

Unknown

Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)

Lanes 1-2 : Merged signal (red and green). Green - ab32081 observed at 41 kDa. Red - loading control ab8245 observed at 37 kDa.

ab32081 Anti-ERK2 antibody [E460] was shown to specifically react with ERK2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265052 (knockout cell lysate ab257525) was used. Wild-type and ERK2 knockout samples were subjected to SDS-PAGE. ab32081 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-ERK2 antibody [E460] (<a href='/en-us/products/primary-antibodies/erk2-antibody-e460-ab32081'>ab32081</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MAPK1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (ab265052)

Predicted band size: 41 kDa

Observed band size: 41 kDa

false

Sanger Sequencing - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)
  • Sanger seq

Unknown

Sanger Sequencing - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)

Allele-2 : Insertion of the selection cassette in exon 2.

Sanger Sequencing - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)
  • Sanger seq

Unknown

Sanger Sequencing - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)

Allele-1 : 1 bp deletion in exon 2.

Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)
  • WB

Lab

Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (AB265052)

All lanes:

Western blot - Anti-ERK2 antibody [E460] (<a href='/en-us/products/primary-antibodies/erk2-antibody-e460-ab32081'>ab32081</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa lysate at 20 µg

Lane 2:

Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (ab265052) at 20 µg

Observed band size: 41 kDa

false

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Immunocytochemistry,Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and Insertion of the selection cassette in exon 2

Disease

Adenocarcinoma

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Complementary Products

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Primary Antibodies

AB184699

Anti-ERK1 + ERK2 antibody [EPR17526]

RabMAb|Recombinant|KO Validated|20ul selling size

Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (AB184699)

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Primary Antibodies

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Anti-p38 alpha/MAPK14 antibody [E229]

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Cell Lines & Lysates

AB257525

Human MAPK1 (ERK2) knockout HeLa cell lysate

Western blot - Human MAPK1 (ERK2) knockout HeLa cell lysate (AB257525)

  • 0
  • 0
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "ICC/IF": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
MAPK1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Immunocytochemistry, Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Supplementary information

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

ERK2 also known as Extracellular signal-Regulated Kinase 2 is a serine/threonine protein kinase in the mitogen-activated protein kinase (MAPK) family with a mass of approximately 42 kDa. This kinase is expressed in many cell types and tissues including the brain liver and lungs. ERK2 plays a significant role in cellular processes such as proliferation differentiation and survival. It is often analyzed using specific assays including ERK2 ELISA and examination of cell lysate samples to determine expression levels and activity.
Biological function summary

ERK2 influences several key cellular functions. It functions as part of a signaling cascade transmitting signals from the exterior to the cell nucleus. In this cascade ERK2 is often part of a multi-protein complex that undergoes sequential phosphorylation. Through these mechanisms ERK2 regulates gene expression and is a pivotal component of the MAPK/ERK pathway ensuring the proper response to growth signals and stress stimuli.

Pathways

ERK2 is a central component of the MAPK/ERK signaling pathway and the PI3K/AKT pathway. These pathways play critical roles in cell cycle regulation and apoptosis. ERK2 activation leads to its interaction with the MEK1/2 proteins which further allows the transmission of mitogenic signals. The interplay between ERK2 and related proteins like E460 often impacts cellular growth and development as it precisely controls the phosphorylation events within the pathway.

ERK2 is connected to certain types of cancer and neurodegenerative diseases. Aberrations in ERK2 signaling pathways are often linked to tumorigenesis where altered interaction with proteins such as Raf and MEK1/2 disrupts cell cycle regulation and apoptosis. In neurodegenerative disorders dysregulated ERK2 activity has been associated with proteins contributing to Alzheimer’s disease indicating its involvement in neuronal survival and stress response mechanisms.
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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

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

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.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

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

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Product protocols

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Scientific reports 15:29838 PubMed40813603

2025

A novel neutrophil extracellular trap-related gene signature for predicting glioblastoma prognosis.

Applications

Unspecified application

Species

Unspecified reactive species

Chao Huang,Xue-Bin Yu,Yong-Zhi Zhou
View all publications
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Product promise

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