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AB265368

Human BCL2L2 knockout HeLa cell line

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BCL2L2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette 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'.
4 Images
Western blot - Human BCL2L2 knockout HeLa cell line (AB265368)
  • WB

Lab

Western blot - Human BCL2L2 knockout HeLa cell line (AB265368)

False colour image of Western blot : Anti-BCL2L2 antibody staining at 1/500 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to BCL2L2. A band was observed at 17 kDa in wild-type HeLa cell lysates with no signal observed at this size in BCL2L2 knockout cell line ab265368 (knockout cell lysate ab258325). To generate this image, wild-type and BCL2L2 knockout HeLa 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 °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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Anti-BCL2L2 antibody at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 40 µg

Lane 2:

Western blot - Human BCL2L2 knockout HeLa cell line (ab265368)

Lane 2:

Western blot - Human BCL2L2 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-bcl2l2-knockout-hela-cell-lysate-ab258325'>ab258325</a>)

Lane 2:

BCL2L2 knockout HeLa cell lysate at 40 µg

Lane 3:

HepG2 cell lysate

Lane 4:

MOLT-4 cell lysate at 40 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Observed band size: 17 kDa

false

Western blot - Human BCL2L2 knockout HeLa cell line (AB265368)
  • WB

Lab

Western blot - Human BCL2L2 knockout HeLa cell line (AB265368)

False colour image of Western blot : Anti-BCL2L2 antibody staining at 1/500 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to BCL2L2. A band was observed at 17 kDa in wild-type HeLa cell lysates with no signal observed at this size in BCL2L2 knockout cell line ab265368 (knockout cell lysate ab258325). To generate this image, wild-type and BCL2L2 knockout HeLa 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 °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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

anti-BCL2L2 antibody at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 40 µg

Lane 2:

BCL2L2 knockout HeLa cell lysate at 40 µg

Lane 2:

Western blot - Human BCL2L2 knockout HeLa cell line (ab265368)

Lane 3:

HepG2 cell lysate at 40 µg

Lane 4:

MOLT-4 cell lysate at 40 µg

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Sanger Sequencing - Human BCL2L2 knockout HeLa cell line (AB265368)
  • Sanger seq

Unknown

Sanger Sequencing - Human BCL2L2 knockout HeLa cell line (AB265368)

Allele-2 : Insertion of the selection cassette in exon 3.

Sanger Sequencing - Human BCL2L2 knockout HeLa cell line (AB265368)
  • Sanger seq

Unknown

Sanger Sequencing - Human BCL2L2 knockout HeLa cell line (AB265368)

Allele-1 : 1 bp insertion in exon 3.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3

Disease

Adenocarcinoma

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Primary Antibodies

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Primary Antibodies

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Cell Lines & Lysates

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Sanger seq": { "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
BCL2L2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
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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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.

Bcl2L2 also known as Bcl-w is a protein that functions as an anti-apoptotic member of the Bcl-2 family. It has a molecular weight of approximately 21 kDa. This protein is expressed in various tissues such as the brain testis and kidney suggesting it plays a significant role in these organs. Bcl2L2's expression pattern and structure contribute to its function in regulating apoptotic pathways by inhibiting cell death signals.
Biological function summary

The function of Bcl-w involves its role in promoting cell survival. It is part of a complex network of interactions within the Bcl-2 family. This protein family includes both pro-apoptotic and anti-apoptotic proteins and Bcl-w helps maintain the balance by impeding apoptosis. The protein localizes to the mitochondrial outer membrane where it sequesters pro-apoptotic members such as Bax and Bak preventing the release of cytochrome c an essential step in the apoptosis process.

Pathways

The Bcl-w protein integrates into the intrinsic mitochondrial apoptosis pathway. It interacts closely with other Bcl-2 family proteins to ensure cellular homeostasis. Important pathways involving Bcl-w include the mitochondrial or intrinsic apoptosis pathway and the PI3K/AKT signaling pathway both of which regulate cell survival. Related proteins include Bcl-2 Bcl-XL Bax and Bak. These proteins collectively modulate the apoptosis signaling and therefore influence cell fate decisions within tissues where Bcl-w is expressed.

Bcl-w has been implicated in cancer and neurodegenerative diseases due to its role in cell survival and apoptosis regulation. In certain cancers the overexpression of Bcl-w protein can lead to unchecked cell proliferation by avoiding apoptosis therefore promoting tumor growth. In neurodegenerative diseases such as Alzheimer's disease altered expression of Bcl-w may contribute to neuronal survival affecting disease progression. Proteins like Bax and Bak which promote apoptosis are often in regulatory interplay with Bcl-w in these conditions highlighting the delicate balance of pro-survival and pro-apoptotic forces in disease states.
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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 BCL2L2
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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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