BCL2L2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.
Apoptosis regulator Bcl-W, B2CL2_HUMAN, BCL 2 Like 2, BCL W, Bcl 2 like 2 protein, Bcl-2-like protein 2, Bcl2-L-2, KIAA0271, PPP1R51, Protein phosphatase 1 regulatory subunit 51
BCL2L2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.
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.
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.
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.
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.
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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False colour image of Western blot: Anti-BCL2L2 antibody staining at 1/500 dilution, shown in green; loading control Anti-alpha Tubulin antibody [DM1A] - 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 Human BCL2L2 knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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
Performed under reducing conditions.
False colour image of Western blot: Anti-BCL2L2 antibody staining at 1/500 dilution, shown in green; loading control Anti-alpha Tubulin antibody [DM1A] - 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 Human BCL2L2 knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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 (Human BCL2L2 knockout HeLa cell lysate ab258325)
Lane 2: BCL2L2 knockout HeLa cell lysate at 40 µg
Lane 3: HepG2 cell lysate
Lane 4: MOLT-4 cell lysate at 40 µg
Lanes 1 - 4: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Lanes 1 - 4: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 17 kDa
Allele-1: 1 bp insertion in exon 3.
Allele-2: Insertion of the selection cassette in exon 3.
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