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    products/cell-lysates/human-bcl10-knockout-hela-cell-lysate-ab257144.pdf

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Cell Biology Apoptosis Intracellular Bcl2 Family
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Human BCL10 knockout HeLa cell lysate (ab257144)

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Western blot - Human BCL10 knockout HeLa cell lysate (ab257144)
  • Western blot - Human BCL10 knockout HeLa cell lysate (ab257144)
  • Sanger Sequencing - Human BCL10 knockout HeLa cell lysate (ab257144)

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Anti-Bcl10 antibody [EPR8587] (ab150380)
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Anti-Bcl10 antibody [EP606Y] (ab33905)

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Overview

  • Product name

    Human BCL10 knockout HeLa cell lysate
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HeLa
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
  • Passage number

    <20
  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    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 - found here. 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.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    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.

    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.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab260892 - Human BCL10 knockout HeLa cell lysate 1 x 100µg
    ab255929 - Human wild-type HeLa cell lysate 1 x 100µg
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Bcl2 Family
    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • CARD Family
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Bcl 2 family
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • NFkB pathway
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Bcl 2 family
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
  • Cell type

    epithelial
  • Disease

    Adenocarcinoma
  • Gender

    Female
  • STR Analysis

    Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10

Target

  • Function

    Promotes apoptosis, pro-caspase-9 maturation and activation of NF-kappa-B via NIK and IKK. May be an adapter protein between upstream TNFR1-TRADD-RIP complex and the downstream NIK-IKK-IKAP complex. Is a substrate for MALT1.
  • Tissue specificity

    Ubiquitous.
  • Involvement in disease

    Note=A chromosomal aberration involving BCL10 is recurrent in low-grade mucosa-associated lymphoid tissue (MALT lymphoma). Translocation t(1;14)(p22;q32). Although the BCL10/IgH translocation leaves the coding region of BCL10 intact, frequent BCL10 mutations could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions.
    Note=Defects in BCL10 are involved in various types of cancer.
  • Sequence similarities

    Contains 1 CARD domain.
  • Post-translational
    modifications

    Phosphorylated. Phosphorylation results in dissociation from TRAF2 and binding to BIRC2/c-IAP2.
  • Cellular localization

    Cytoplasm > perinuclear region. Membrane raft. Appears to have a perinuclear, compact and filamentous pattern of expression. Also found in the nucleus of several types of tumor cells. Colocalized with DPP4 in membrane rafts.
  • Target information above from: UniProt accession O95999 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • AI132454
    • B cell CLL/lymphoma 10
    • B cell lymphoma/leukemia10
    • B-cell CLL/lymphoma 10
    • B-cell leukemia/lymphoma 10
    • B-cell lymphoma/leukemia 10
    • Bcl 10
    • Bcl-10
    • Bcl10
    • BCL10_HUMAN
    • c E10
    • c-E10
    • C81403
    • CARD containing apoptotic signaling protein
    • CARD containing molecule enhancing NF kappa B
    • CARD containing molecule enhancing NF kB
    • CARD containing molecule enhancing NF-kB
    • CARD containing molecule enhancing NFkB
    • CARD containing proapoptotic protein
    • CARD like apoptotic protein
    • CARD-containing apoptotic signaling protein
    • CARD-containing molecule enhancing NF-kappa-B
    • CARD-containing proapoptotic protein
    • CARD-like apoptotic protein
    • CARMEN
    • Caspase recruiting domain containing protein
    • caspase-recruiting domain-containing protein
    • cCARMEN
    • cE 10
    • cE10
    • CED 3/ICH 1 prodomain homologous E10 like regulator
    • CED-3/ICH-1 prodomain homologous E10-like regulator
    • CED3/ICH1 prodomain homologous E10 like regulator
    • Cellular E10
    • Cellular homolog of vCARMEN
    • Cellular-E10
    • CIPER
    • CLAP
    • hCLAP
    • Mammalian CARD containing adapter molecule E10
    • Mammalian CARD-containing adapter molecule E10
    • mE 10
    • mE10
    • R-RCD1
    see all

Associated products

  • KO cell lines

    • Human BCL10 knockout HeLa cell line (ab261797)
  • Related Products

    • Anti-Bcl10 antibody [EPR8587] (ab150380)
    • Anti-Bcl10 antibody [EP606Y] - BSA and Azide free (ab189219)
    • Anti-Bcl10 antibody [EPR8587] - BSA and Azide free (ab248953)
    • Anti-Bcl10 antibody [EP606Y] (ab33905)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab257144 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 26 kDa.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 26 kDa.

Images

  • Western blot - Human BCL10 knockout HeLa cell lysate (ab257144)
    Western blot - Human BCL10 knockout HeLa cell lysate (ab257144)

    Lane 1: Wild-type HeLa cell lysate (20 µg)

    Lane 2: BCL10 knockout HeLa cell lysate (20 µg)

    Lane 3: Ramos cell lysate (20 µg)

    Lane 4: A549 cell lysate (20 µg)

    Lanes 1-4: Merged signal (red and green). Green - ab150380 observed at 32 kDa. Red - loading control, ab7291 observed at 52 kDa.

    ab150380 Anti-Bcl10 antibody [EPR8587] was shown to specifically react with Bcl10 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261797 (knockout cell lysate ab257144) was used. Wild-type and Bcl10 knockout samples were subjected to SDS-PAGE. ab150380 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Human BCL10 knockout HeLa cell lysate (ab257144)
    Western blot - Human BCL10 knockout HeLa cell lysate (ab257144)

    Lane 1: Wild-type HeLa cell lysate (20 µg)

    Lane 2: BCL10 knockout HeLa cell lysate (20 µg)

    Lane 3: Ramos cell lysate (20 µg)

    Lane 4: A549 cell lysate (20 µg)

    Lanes 1-4: Merged signal (red and green). Green - ab33905 observed at 32 kDa. Red - loading control, ab7291 observed at 52 kDa.

    ab33905 Anti-Bcl10 antibody [EP606Y] was shown to specifically react with Bcl10 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261797 (knockout cell lysate ab257144) was used. Wild-type and Bcl10 knockout samples were subjected to SDS-PAGE. ab33905 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.

  • Sanger Sequencing - Human BCL10 knockout HeLa cell lysate (ab257144)
    Sanger Sequencing - Human BCL10 knockout HeLa cell lysate (ab257144)
    Homozygous: 1 bp insertion in exon 1

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab257144? Please let us know so that we can cite the reference in this datasheet.

ab257144 has not yet been referenced specifically in any publications.

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