Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [31/PBK] to PBK/SPK - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free
    See all PBK/SPK primary antibodies

  • Description

    Mouse monoclonal [31/PBK] to PBK/SPK - BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HEK-293T whole cell lysate. HeLa, HepG2, RAW264.7, C6, PC-12 and U-87 MG whole cell lysate. Human, rat and mouse testis tissue lysate. IHC-P: Human tonsil tissue. Mouse and rat testis tissue. Flow Cyt (intra): HEK-239T cells.

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    ab280216 is the carrier-free version of ab280209. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab280216 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    Constituent: 100% PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    31/PBK

  • Isotype

    IgG2b

Images

  • Western blot - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    Western blot - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    All lanes : Anti-PBK/SPK antibody [31/PBK] (ab280209) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
    Lane 2 : PBK knockout HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
    Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
    Lane 4 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution

    Predicted band size: 36 kDa



    This image was produced using ab280209, the same clone in a different formulation.

    Lanes 1 - 4: Merged signal (red and green). Green - ab280209 observed at 36 kDa. Red - loading control ab52901(Rabbit monoclonal [EP1331Y] to beta Tubulin) observed at 52 kDa.

    Lanes 1-2: ab280209 Anti-PBK antibody was shown to react with in HEK-293T cells in Western blot. Loss of signal was observed when PBK knockout sample was used. Wild-type and PBK knockout samples were subjected to SDS-PAGE.

    ab280209 and Anti-beta Tubulin antibody [EP1331Y] (ab52901) were incubated at 4° overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

    Blocking/Dilution buffer: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)

    This image was produced using ab280209, the same clone in a different formulation.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PBK/PSK with ab280209 at 1/500 (1.946 µg/ml) dilution followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Positive staining in germinal center of human tonsil.

    The section was incubated with ab280209 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Flow Cytometry - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    Flow Cytometry - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)

    This image was produced using ab280209, the same clone in a different formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling PBK/SPK with ab280209 at 1/1000 dilution (0.1 µg) (red) compared with a mouse monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    A Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

  • Western blot - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    Western blot - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    All lanes : Anti-PBK/SPK antibody [31/PBK] (ab280209) at 1/1000 dilution

    Lane 1 : Human testis tissue lysate
    Lane 2 : Human heart tissue lysate
    Lane 3 : Mouse testis tissue lysate
    Lane 4 : Mouse brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 36 kDa



    This image was produced using ab280209, the same clone in a different formulation.

    Negative control: Human heart, mouse brain (PMID: 27898655).

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:23547718).

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    Western blot - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    All lanes : Anti-PBK/SPK antibody [31/PBK] (ab280209) at 1/1000 dilution

    Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
    Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
    Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
    Lane 4 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
    Lane 5 : C6 (rat glial tumor glial cell), whole cell lysate
    Lane 6 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
    Lane 7 : Rat testis tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 36 kDa



    This image was produced using ab280209, the same clone in a different formulation.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:23547718).

    Exposure time: lane 1: 37 secs; lane 2-7: 15 secs.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)

    This image was produced using ab280209, the same clone in a different formulation.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling PBK/PSK with ab280209 at 1/500 (1.946 µg/ml) dilution followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Positive staining in mouse testis.

    The section was incubated with ab280209 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)

    This image was produced using ab280209, the same clone in a different formulation.

    Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling PBK/PSK with ab280209 at 1/500 (1.946 µg/ml) dilution followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Positive staining in rat testis.

    The section was incubated with ab280209 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)
    Anti-PBK/SPK antibody [31/PBK] - BSA and Azide free (ab280216)

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