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AB265708

Human SLC3A2 (CD98) knockout HeLa cell line

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SLC3A2 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 6. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
5 Images
Immunocytochemistry/ Immunofluorescence - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SLC3A2 (CD98) KO HeLa (SLC3A2 (CD98) knockout human cervix adenocarcinoma epithelial cell) (ab265708) cells labelling CD98 with ab307587 at 1/100 (4.97 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green). Confocal image showing membrane and weak cytoplasmic staining in parental HeLa cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

Western blot - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)
  • WB

Lab

Western blot - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)

Lanes 1 - 5 : Merged signal (red and green). Green - ab253273 observed at 80 - 90 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

ab253273 was shown to react with CD98 in wild-type HeLa cells in Western blot with loss of signal observed in SLC3A2 knockout cell line ab265708 (SLC3A2 knockout cell lysate ab257680). Wild-type HeLa and SLC3A2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween®) before incubation with ab253273 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Human SLC3A2 (CD98) knockout HeLa cell line (ab265708)

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Sanger Sequencing - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)
  • Sanger seq

Unknown

Sanger Sequencing - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)

Homozygous : 14 bp deletion in exon 6.

Immunocytochemistry/ Immunofluorescence - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SLC3A2 (CD98) KO HeLa (SLC3A2 (CD98) knockout human cervix adenocarcinoma epithelial cell) (ab265708) cells labelling CD98 with ab314339 at 1/50 (10.0 ug/ml) dilution(Red). Confocal image showing membrane staining in parental HeLa cell line, while no staining in SLC3A2 knockout HeLa cells (ab265708). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195887 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 488) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Green). The Nuclear counterstain was DAPI (Blue).

Immunocytochemistry/ Immunofluorescence - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human SLC3A2 (CD98) knockout HeLa cell line (AB265708)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SLC3A2 (CD98) KO HeLa (SLC3A2 (CD98) knockout human cervix adenocarcinoma epithelial cell) (ab265708) cells labelling CD98 with ab314338 at 1/50 (10.0 ug/ml) dilution(Red). Confocal image showing membrane staining in parental HeLa cell line, while no staining in SLC3A2 knockout HeLa cells (ab265708). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195887 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 488) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Green). The Nuclear counterstain was DAPI (Blue).

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 6

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB307587

Anti-CD98 antibody [EPR27110-42]

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] (AB307587)

  • 0
  • 0
Primary Antibodies

AB208776

Anti-SLC7A5/LAT1 antibody [EPR17573]

RabMAb|Recombinant|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR17573] (AB208776)

  • 0
  • 0
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Primary Antibodies

AB8227

Anti-beta Actin antibody - Loading Control

BOND RX™ Validated|Lab Essentials

Western blot - Anti-beta Actin antibody - Loading Control (AB8227)

  • 5
  • 104
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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
SLC3A2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
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 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.
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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Supplementary information

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

CD98 also known as CD98hc CD98 protein or LAT1/CD98 is a type II membrane protein often found in association with the heavy chains of amino acid transporters like LAT1. It has a molecular mass of approximately 85-100 kDa. CD98 is widely expressed in numerous tissues including the liver kidney and various tumors. It plays an important role in cellular processes by facilitating the transport of amino acids across plasma membranes which is essential for maintaining cellular homeostasis and proliferation.
Biological function summary

CD98 forms a complex with light chains of the heterodimeric amino acid transporters such as LAT1 and LAT2. This interaction enhances the uptake of neutral amino acids into cells. CD98 also interacts with integrins influencing cell adhesion and migration which are critical for cellular signaling and response. Through these interactions CD98 impacts not only cell survival and growth but also immune responses and angiogenesis.

Pathways

CD98 is a vital component of the mTOR signaling pathway influencing cell growth and proliferation. It also plays a part in the integrin signaling pathway which regulates cell adhesion and migration. In these pathways CD98 works closely with proteins like FG1 and mTORC1. These relationships enable it to modulate cellular responses to nutritional availability and mechanical cues from the environment.

CD98 expression and function have links to cancer and autoimmune diseases. High expression levels of CD98 are observed in various cancers such as lymphomas where it correlates with increased tumor growth and poor prognosis. In autoimmune disorders like rheumatoid arthritis CD98 influences immune cell activation and migration. Proteins such as CP1 and FG1 interact with CD98 contributing to disease pathology by affecting cellular and immune responses.
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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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Target data

See full target information SLC3A2
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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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