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AB270472

Human CBFB knockout A-431 cell line

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CBFB KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 19 bp deletion Frameshift = 99.99%.

View Alternative Names

CBF b, CBF-beta, Core binding factor beta subunit, Core-binding factor subunit beta, Core-binding factor, beta subunit (CBFB), transcript variant 2, PEA 2, PEA2-beta, PEBB_HUMAN, PEBP 2B, PEBP2-beta, Polyomavirus enhancer-binding protein 2 beta subunit, SL3 3 enhancer factor 1 beta subunit, SL3-3 enhancer factor 1 subunit beta, SL3/AKV core-binding factor beta subunit

4 Images
Western blot - Human CBFB knockout A-431 cell line (AB270472)
  • WB

Lab

Western blot - Human CBFB knockout A-431 cell line (AB270472)

Lanes 1 - 4 : Merged signal (red and green). Green - ab124693 observed at 24-25 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab124693 was shown to react with CBFb in wild-type A-431 cells in western blot Loss of signal was observed when CBFB knockout cell line ab270472 (knockout cell lysate ab270495) was used. Wild-type and CBFB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab124693 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CBFb antibody [EPR6321] (<a href='/en-us/products/primary-antibodies/cbfb-antibody-epr6321-ab124693'>ab124693</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

CBFB knockout A431 cell lysate at 20 µg

Lane 2:

Western blot - Human CBFB knockout A-431 cell line (ab270472)

Lane 3:

K562 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 24-25 kDa

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Western blot - Human CBFB knockout A-431 cell line (AB270472)
  • WB

Lab

Western blot - Human CBFB knockout A-431 cell line (AB270472)

ab133600 was shown to react with CBFb in wild-type A-431 cells in western blot. Loss of signal was observed when CBFB knockout cell line ab270472 (knockout cell lysate ab270495) was used. Membranes were blocked in 2 % BSA in TBS-T (0.1 % Tween®) before incubation with ab133600 overnight at 4°C at a 1 in 1000 dilution and ab184095 (Mouse Anti-GAPDH antibody [mAbcam 9484] - Alexa Fluor® 680) at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1/5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.

All lanes:

Western blot - Anti-CBFb antibody [EPR6322] (<a href='/en-us/products/primary-antibodies/cbfb-antibody-epr6322-ab133600'>ab133600</a>) at 1/1000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

CBFB knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CBFB knockout A-431 cell line (ab270472)

Lane 3:

K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 24 kDa

false

Exposure time: 150s

Western blot - Human CBFB knockout A-431 cell line (AB270472)
  • WB

Lab

Western blot - Human CBFB knockout A-431 cell line (AB270472)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133600 observed at 22 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab133600 was shown to react with CBFb in wild-type A-431 cells in western blot with loss of signal observed in CBFB knockout cell line ab270472 (knockout cell lysate ab270495). Wild-type and CBFB knockout A-431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 2% BSA in TBS-T (0.1% Tween®) before incubation with ab133600 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CBFb antibody [EPR6322] (<a href='/en-us/products/primary-antibodies/cbfb-antibody-epr6322-ab133600'>ab133600</a>) at 1/1000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

CBFB knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CBFB knockout A-431 cell line (ab270472)

Lane 3:

K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 22 kDa

false

Next Generation Sequencing - Human CBFB knockout A-431 cell line (AB270472)
  • NGS

Supplier Data

Next Generation Sequencing - Human CBFB knockout A-431 cell line (AB270472)

Knockout achieved by CRISPR/Cas9; X = 19 bp deletion; Frameshift = 99.99%

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 19 bp deletion Frameshift = 99.99%

Disease

Epidermoid Carcinoma

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Reactivity data

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Product details

Recommended control: Human wild-type A-431 cell line (ab263975). 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 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
CBFB
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation 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.

CBFb also known as Core-Binding Factor Subunit Beta weighs approximately 22 kDa. It plays an important role in gene regulation by stabilizing the DNA binding of core-binding factor alpha (CBFA) subunits such as RUNX proteins. CBFb does not bind DNA directly. Instead it enhances the affinity of the RUNX family for their DNA targets. CBFb is broadly expressed in various tissues with high levels observed in hematopoietic cells.
Biological function summary

CBFb forms a complex with RUNX transcription factors facilitating their function. The complex is essential for hematopoiesis as it regulates the expression of genes critical for blood cell differentiation and proliferation. CBFb is involved in bone development influencing the formation of bone-related genes. This CBFb/RUNX complex plays a role in regulating the cell cycle and apoptosis fundamental processes for maintaining cellular integrity.

Pathways

More than one signaling pathway includes CBFb's involvement. Particularly it participates in the Wnt signaling pathway an important player in cell differentiation. Additionally CBFb/RUNX complex regulates genes downstream in the TGF-beta signaling pathway impacting cellular proliferation and differentiation. Through these pathways CBFb is linked to proteins like SMAD3 further emphasizing its role in mediating various biological processes.

Abnormalities in CBFb function connect to acute myeloid leukemia (AML). CBFb's interaction with RUNX1 can result in a chromosomal translocation that generates the CBFb-MYH11 fusion protein which disrupts normal hematopoiesis and contributes to leukemogenesis. Moreover CBFb variations associate with cleidocranial dysplasia a skeletal disorder through its interaction with the RUNX2 protein highlighting its significance in bone development.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

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AB133600

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Western blot - Anti-CBFb antibody [EPR6322] (AB133600)

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Cell Lines & Lysates

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Western blot - Human CBFB knockout A-431 cell lysate (AB270495)

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Proteins & Peptides

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