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AB255363

Human BAX knockout HeLa cell line

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BAX 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 deletion in exon 2 and Insertion of the selection cassette in exon 2.

View Alternative Names

Apoptosis regulator BAX, BAXA, BAX_HUMAN, BCL2 associated X protein, BCL2 associated X protein omega, BCL2 associated X protein transcript variant delta2, Bax-protein, Baxdelta2G9, Baxdelta2G9omega, Baxdelta2omega, Bcl-2-like protein 4, Bcl2-L-4, membrane isoform alpha

4 Images
Western blot - Human BAX knockout HeLa cell line (AB255363)
  • WB

Lab

Western blot - Human BAX knockout HeLa cell line (AB255363)

Lanes 1- 2 : Merged signal (red and green). Green - ab182734 observed at 21 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab182734 was shown to react with Bax in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255363 (knockout cell lysate ab263841) was used. Wild-type HeLa and BAX knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab182734 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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-Bax antibody [EPR18284] (<a href='/en-us/products/primary-antibodies/bax-antibody-epr18284-ab182734'>ab182734</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BAX knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BAX knockout HeLa cell line (ab255363)

Predicted band size: 21 kDa

Observed band size: 21 kDa

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Western blot - Human BAX knockout HeLa cell line (AB255363)
  • WB

Lab

Western blot - Human BAX knockout HeLa cell line (AB255363)

Lanes 1- 2 : Merged signal (red and green). Green - ab32503. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32503 was shown to react with Bax in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255363 (knockout cell lysate ab263841) was used. Wild-type HeLa and BAX knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32503 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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-Bax antibody [E63] (<a href='/en-us/products/primary-antibodies/bax-antibody-e63-ab32503'>ab32503</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BAX knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-bax-knockout-hela-cell-lysate-ab263841'>ab263841</a>) at 20 µg

Lane 2:

Western blot - Human BAX knockout HeLa cell line (ab255363)

Predicted band size: 21 kDa

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Sanger Sequencing - Human BAX knockout HeLa cell line (AB255363)
  • Sanger seq

Unknown

Sanger Sequencing - Human BAX knockout HeLa cell line (AB255363)

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

Sanger Sequencing - Human BAX knockout HeLa cell line (AB255363)
  • Sanger seq

Unknown

Sanger Sequencing - Human BAX knockout HeLa cell line (AB255363)

Allele-1 : 1 bp deletion in exon 2.

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 deletion in exon 2 and Insertion of the selection cassette in exon 2

Disease

Adenocarcinoma

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Reactivity data

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Product details

Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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
BAX
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.

Bax also known as Bcl-2-associated X protein plays an important role in the regulation of apoptosis. It is widely known for its pro-apoptotic function opposing the action of anti-apoptotic proteins like Bcl-2. Bax has a molecular weight of approximately 21 kDa making it identifiable in laboratory techniques such as western blot. Researchers often use anti-Bax antibodies to detect and analyze its expression levels in various studies. The Bax protein is mainly expressed in mitochondria where it undergoes changes in response to apoptotic signals.
Biological function summary

The Bax protein forms homodimers or heterodimers with other Bcl-2 family proteins to facilitate apoptosis. This protein is part of a larger complex involving several members of the Bcl-2 family which finely tunes the balance between cell survival and programmed cell death. Through a change in its conformation Bax translocates to the outer mitochondrial membrane where it promotes the release of cytochrome c and other apoptogenic factors into the cytosol.

Pathways

Bax plays a significant role in the intrinsic pathway of apoptosis also known as the mitochondrial pathway. This pathway involves the release of cytochrome c which further activates downstream proteins such as caspase-9 and caspase-3. Bax interacts closely with proteins like Bak another pro-apoptotic member of the Bcl-2 family providing a cooperative function in mitochondrial membrane permeabilization. Together Bax and Bak act as gatekeepers of the mitochondrial pathway determining the cell's fate in response to apoptotic stimuli.

The dysregulation of Bax often associates with various cancers and neurodegenerative diseases. In cancer reduced Bax expression or function can lead to resistance to apoptosis contributing to unchecked cellular proliferation. Conversely in neurodegenerative disorders such as Alzheimer's disease increased Bax activity results in excessive neuronal apoptosis. The balance between Bax and its partner proteins like Bcl-2 or Bcl-xL is important in maintaining normal cellular function and preventing disease progression.
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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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