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  1. Link

    human-myc-c-myc-knockout-hek-293t-cell-lysate-ab263850.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families HLH / Leucine Zipper HLH / Leucine Zipper
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Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)

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Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
  • Western blot - Human MYC knockout HEK293T cell lysate (ab263850)
  • Sanger Sequencing - Human MYC knockout HEK293T cell lysate (ab263850)

You may also be interested in

Peptide
Human c-Myc peptide (ab166837)
Knockout
Product image
Human MYC (c-Myc) knockout HEK-293T cell line (ab256500)
Primary
Product image
Anti-c-Myc antibody [Y69] - ChIP Grade (ab32072)

View more associated products

Overview

  • Product name

    Human MYC (c-Myc) knockout HEK-293T cell lysate
    See all c-Myc kits
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HEK293T
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous (4N): 1bp T insertion (2N); 8 bp deletion and C to T insertion (1N); 4 bp deletion in exon 2 (1N)
  • 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
    ab256769 - Human MYC knockout HEK293T cell lysate 1 x 100µg
    ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • HLH / Leucine Zipper
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Nuclear
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Cell cycle
    • Cell differentiation
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Transcription factors
    • Cancer
    • Tumor biomarkers
    • Oncoproteins
  • Cell type

    epithelial
  • STR Analysis

    Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12

Target

  • Function

    Participates in the regulation of gene transcription. Binds DNA in a non-specific manner, yet also specifically recognizes the core sequence 5'-CAC[GA]TG-3'. Seems to activate the transcription of growth-related genes.
  • Involvement in disease

    Note=Overexpression of MYC is implicated in the etiology of a variety of hematopoietic tumors.
    Note=A chromosomal aberration involving MYC may be a cause of a form of B-cell chronic lymphocytic leukemia. Translocation t(8;12)(q24;q22) with BTG1.
    Defects in MYC are a cause of Burkitt lymphoma (BL) [MIM:113970]. A form of undifferentiated malignant lymphoma commonly manifested as a large osteolytic lesion in the jaw or as an abdominal mass. Note=Chromosomal aberrations involving MYC are usually found in Burkitt lymphoma. Translocations t(8;14), t(8;22) or t(2;8) which juxtapose MYC to one of the heavy or light chain immunoglobulin gene loci.
  • Sequence similarities

    Contains 1 basic helix-loop-helix (bHLH) domain.
  • Post-translational
    modifications

    Phosphorylated by PRKDC. Phosphorylation at Thr-58 and Ser-62 by GSK3 is required for ubiquitination and degradation by the proteasome.
    Ubiquitinated by the SCF(FBXW7) complex when phosphorylated at Thr-58 and Ser-62, leading to its degradation by the proteasome. In the nucleoplasm, ubiquitination is counteracted by USP28, which interacts with isoform 1 of FBXW7 (FBW7alpha), leading to its deubiquitination and preventing degradation. In the nucleolus, however, ubiquitination is not counteracted by USP28, due to the lack of interaction between isoform 4 of FBXW7 (FBW7gamma) and USP28, explaining the selective MYC degradation in the nucleolus. Also polyubiquitinated by the DCX(TRUSS) complex.
  • Cellular localization

    Nucleus > nucleoplasm. Nucleus > nucleolus.
  • Target information above from: UniProt accession P01106 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Form

    c-Myc is also expressed in the cytoplasm.
  • Alternative names

    • AU016757
    • Avian myelocytomatosis viral oncogene homolog
    • bHLHe39
    • c Myc
    • Cellular myelocytomatosis oncogene
    • Class E basic helix-loop-helix protein 39
    • MGC105490
    • MRTL
    • Myc
    • Myc protein
    • Myc proto oncogene protein
    • Myc proto-oncogene protein
    • myc-related translation/localization regulatory factor
    • MYC_HUMAN
    • Myc2
    • myca
    • MYCC
    • Myelocytomatosis oncogene
    • Myelocytomatosis oncogene a
    • Niard
    • Nird
    • oncogene c-Myc
    • Oncogene Myc
    • OTTHUMP00000158589
    • OTTHUMP00000227763
    • Proto-oncogene c-Myc
    • Protooncogene homologous to myelocytomatosis virus
    • RNCMYC
    • Transcription factor p64
    • Transcriptional regulator Myc-A
    • V-Myc avian myelocytomatosis viral oncogene homolog
    • v-myc myelocytomatosis viral oncogene homolog (avian)
    • zc-myc
    see all

Associated products

  • KO cell lines

    • Human MYC (c-Myc) knockout HEK-293T cell line (ab256500)
  • KO cell pellets

    • Human MYC (c-Myc) knockout HEK-293T cell pellet (ab278810)
  • Related Products

    • Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)
    • Anti-c-Myc antibody [Y69] - ChIP Grade (ab32072)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab263850 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.
Notes
WB
Use at an assay dependent concentration.

Images

  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
    Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)

    Lane 1: Wild-type HEK-293T cell lysate 20 μg
    Lane 2: MYC knockout HEK-293T cell lysate 20 μg
    Lane 3: Jurkat cell lysate 20 μg
    Lane 4: SH-SY5Y cell lysate 20 μg
    False colour image of Western blot: Anti-Myc tag antibody [Myc.A7] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab18185 was shown to bind specifically to Myc tag. A band was observed at 57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC knockout cell line ab256500 (knockout cell lysate ab263850). The band observed in the knockout lysate lane below 57 kDa is likely to represent a truncated form of Myc tag. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC knockout HEK-293T cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
    Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)

    Lane 1: Wild-type HEK-293T cell lysate 20 μg
    Lane 2: MYC CRISPR-Cas9 edited HEK-293T cell lysate 20 μg
    Lane 3: Jurkat cell lysate 20 μg
    Lane 4: SH-SY5Y cell lysate 20 μg
    False colour image of Western blot: Anti-Myc tag antibody [Myc.A7] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab18185 was shown to bind specifically to Myc tag. A band was observed at 57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC CRISPR-Cas9 edited cell line ab256500 (CRISPR-Cas9 edited cell lysate ab263850). The band observed in the CRISPR-Cas9 edited lysate lane below 57 kDa is likely to represent a truncated form of Myc tag. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC CRISPR-Cas9 edited HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
    Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)

    Lane 1: Wild-type HEK-293T cell lysate 20 μg
    Lane 2: MYC knockout HEK-293T cell lysate 20 μg
    Lane 3: Jurkat cell lysate 20 μg
    Lane 4: SH-SY5Y cell lysate 20 μg
    False colour image of Western blot: Anti-Myc tag antibody [9E10] staining at 1/200 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32 was shown to bind specifically to Myc tag. A band was observed at 57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC knockout cell line ab256500 (knockout cell lysate ab263850). The band observed in the knockout lysate lane below 57 kDa is likely to represent a truncated form of Myc tag. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC knockout HEK-293T cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
    Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)

    Lane 1: Wild-type HEK-293T cell lysate 20 μg
    Lane 2: MYC CRISPR-Cas9 edited HEK-293T cell lysate 20 μg
    Lane 3: Jurkat cell lysate 20 μg
    Lane 4: SH-SY5Y cell lysate 20 μg
    False colour image of Western blot: Anti-Myc tag antibody [9E10] staining at 1/200 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32 was shown to bind specifically to Myc tag. A band was observed at 57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC CRISPR-Cas9 edited cell line ab256500 (CRISPR-Cas9 edited cell lysate ab263850). The band observed in the CRISPR-Cas9 edited lysate lane below 57 kDa is likely to represent a truncated form of Myc tag. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC CRISPR-Cas9 edited HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
    Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)

    Lane 1: Wild-type HEK-293T cell lysate 20 μg
    Lane 2: MYC knockout HEK-293T cell lysate 20 μg
    Lane 3: Jurkat cell lysate 20 μg
    Lane 4: SH-SY5Y cell lysate 20 μg
    False colour image of Western blot: Anti-c-Myc antibody [Y69] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32072 was shown to bind specifically to c-Myc. A band was observed at 45/57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC knockout cell line ab256500 (knockout cell lysate ab263850). The band observed in the knockout lysate lane below 45/57 kDa is likely to represent a truncated form of c-Myc. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC knockout HEK-293T cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
    Western blot - Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)

    Lane 1: Wild-type HEK-293T cell lysate 20 μg
    Lane 2: MYC CRISPR-Cas9 edited HEK-293T cell lysate 20 μg
    Lane 3: Jurkat cell lysate 20 μg
    Lane 4: SH-SY5Y cell lysate 20 μg
    False colour image of Western blot: Anti-c-Myc antibody [Y69] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32072 was shown to bind specifically to c-Myc. A band was observed at 45/57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC CRISPR-Cas9 edited cell line ab256500 (CRISPR-Cas9 edited cell lysate ab263850). The band observed in the CRISPR-Cas9 edited lysate lane below 45/57 kDa is likely to represent a truncated form of c-Myc. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC CRISPR-Cas9 edited HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Human MYC knockout HEK293T cell lysate (ab263850)
    Western blot - Human MYC knockout HEK293T cell lysate (ab263850)

    Lane 1: Jurkat cell lysate (20 µg)

    Lane 2: HeLa cell lysate (20 µg)

    Lane 3: Wild-type HEK-293T cell lysate (20 µg)

    Lane 4: MYC knockout HEK-293T cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32072 observed at 57 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab32072 was shown to react with MYC in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab256500 (knockout cell lysate ab263850) was used. Wild-type and MYC knockout samples were subjected to SDS-PAGE. ab32072 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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 20000 dilution for 1 hour at room temperature before imaging.

  • Sanger Sequencing - Human MYC knockout HEK293T cell lysate (ab263850)
    Sanger Sequencing - Human MYC knockout HEK293T cell lysate (ab263850)
    Homozygous: 1 bp insertion in exon 2

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 ab263850? Please let us know so that we can cite the reference in this datasheet.

ab263850 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Used as control for Myc antibody, compared to normal Hek293T lysate. Worked well.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Nadeem Shaikh

Verified customer

Submitted May 12 2021

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