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AB270495

Human CBFB knockout A-431 cell lysate

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CBFB KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. 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 lysate (AB270495)
  • WB

Lab

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

Lane 1 : Wild-type A431 cell lysate 20 ug

Lane 2 : CBFB knockout A431 cell lysate 20 ug

Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate 20 ug

Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

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 (<a href='/en-us/products/cell-lines/human-cbfb-knockout-a-431-cell-line-ab270472'>ab270472</a>)

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 lysate (AB270495)
  • WB

Lab

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

Lane 1 : Wild-type A431 cell lysate 20 ug

Lane 2 : CBFB knockout A431 cell lysate 20 ug

Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate 20 ug

Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

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 (<a href='/en-us/products/cell-lines/human-cbfb-knockout-a-431-cell-line-ab270472'>ab270472</a>)

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 lysate (AB270495)
  • WB

Lab

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

Lane 1 : Wild-type A431 cell lysate 20 ug

Lane 2 : CBFB knockout A431 cell lysate 20 ug

Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate 20 ug

Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

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 (<a href='/en-us/products/cell-lines/human-cbfb-knockout-a-431-cell-line-ab270472'>ab270472</a>)

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 lysate (AB270495)
  • NGS

Supplier Data

Next Generation Sequencing - Human CBFB knockout A-431 cell lysate (AB270495)

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

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

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

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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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