Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Creatine kinase B type antibody [EPR3927] (ab92452)

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Overview

  • Product name

    Anti-Creatine kinase B type antibody [EPR3927]
    See all Creatine kinase B type primary antibodies

  • Description

    Rabbit monoclonal [EPR3927] to Creatine kinase B type

  • Host species

    Rabbit

  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Creatine kinase B type aa 350-450 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, HAP1, SHSY-5Y, Y-79, SW840, mouse cerebellum, mouse brain, C6 and rat brain whole cell lysates IHC-P: Human astrocytoma and glioma tissue ICC/IF: HeLa cells FC: HeLa cells

  • General notes

     

     

    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.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab92452 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 1/10000 - 1/50000. Predicted molecular weight: 43 kDa.
IHC-P 1/100 - 1/1000.

See IHC antigen retrieval protocols.

Use of an HRP/AP polymerized secondary antibody is recommended for enhanced staining.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Perform heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
ICC/IF 1/50 - 1/500.

Target

  • Function

    Reversibly catalyzes the transfer of phosphate between ATP and various phosphogens (e.g. creatine phosphate). Creatine kinase isoenzymes play a central role in energy transduction in tissues with large, fluctuating energy demands, such as skeletal muscle, heart, brain and spermatozoa.

  • Sequence similarities

    Belongs to the ATP:guanido phosphotransferase family.
    Contains 1 phosphagen kinase C-terminal domain.
    Contains 1 phosphagen kinase N-terminal domain.

  • Cellular localization

    Cytoplasm.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • B CK antibody
    • B-CK antibody
    • BB-CK antibody
    • BCK antibody
    • Brain creatine kinase antibody
    • Ckb antibody
    • CKBB antibody
    • Creatine kinase B antibody
    • Creatine kinase B chain antibody
    • Creatine kinase B type antibody
    • Creatine kinase B-type antibody
    • Creatine Kinase BB Isoenzyme antibody
    • Creatine kinase brain antibody
    • Creatine kinase brain type antibody
    • Creatine phosphokinase BB antibody
    • Epididymis luminal protein 211 antibody
    • Epididymis secretory protein Li 29 antibody
    • HEL 211 antibody
    • HEL S 29 antibody
    • KCRB_HUMAN antibody
    see all

Images

  • Western blot - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Western blot - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Creatine kinase B knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: SH-SY5Y cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab92452 observed at 45 kDa. Red - loading control, ab18058, observed at 2X kDa.

    ab92452 (unpurified) was shown to specifically react with Creatine kinase B when ProteinX knockout samples were used. Wild-type and Creatine kinase B knockout samples were subjected to SDS-PAGE. ab92452 and ab18058 (loading control to vinculin) were both diluted 1/10 000 and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Western blot - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Anti-Creatine kinase B type antibody [EPR3927] (ab92452) at 1/10000 dilution (Purified) + Y79 (Human retinoblastoma retinoblastoma) whole cell lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa
    Observed band size: 43 kDa

  • Western blot - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Western blot - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Lanes 1-4 : Anti-Creatine kinase B type antibody [EPR3927] (ab92452) at 1/10000 dilution (Purified)
    Lane 5 : Anti-Creatine kinase B type antibody [EPR3927] (ab92452) at 1/10000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse cerebellum lysates
    Lane 3 : Mouse brain lysates
    Lane 4 : C6 (Rat glial tumor glial cell) whole cell lysates
    Lane 5 : Rat brain lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa
    Observed band size: 43 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human astrocytoma tissue sections labeling Creatine kinase B type with purified ab92452 at 1/1000 dilution (0.101 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Immunocytochemistry/ Immunofluorescence - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Creatine kinase B type with purified ab92452 at 1:50 dilution (2.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Flow Cytometry - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Creatine kinase B type with purified ab92452 at 1/20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Western blot - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Western blot - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    All lanes : Anti-Creatine kinase B type antibody [EPR3927] (ab92452) at 1/10000 dilution (unpurified)

    Lane 1 : SHSY-5Y cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : Y-79 cell lysate
    Lane 4 : SW840 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 43 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)

    Paraffin embedded Human glioma tissue labelled with ab92452 (unpurified) at 1/100 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Immunocytochemistry/ Immunofluorescence - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)

    Immunofluorescence analysis of HeLa cells labelled with ab92452 (unpurified) at 1/100 dilution.

  • Flow Cytometry - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)
    Flow Cytometry - Anti-Creatine kinase B type antibody [EPR3927] (ab92452)

    Overlay histogram showing HeLa cells stained with ab92452 (unpurified) (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92452, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

ab92452 has not yet been referenced specifically in any publications.

Publishing research using ab92452? Please let us know so that we can cite the reference in this datasheet.

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