Validated using a knockout cell line
Recombinant
RabMAb

Anti-KIF5B antibody [EPR10276(B)] (ab167429)

Overview

  • Product name
    Anti-KIF5B antibody [EPR10276(B)]
    See all KIF5B primary antibodies
  • Description
    Rabbit monoclonal [EPR10276(B)] to KIF5B
  • 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, ICC/IF, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human KIF5B aa 1-100.
    Database link: P33176

  • Positive control
    • Human fetal kidney, HeLa, Jurkat and HepG2 lysates; Human brain tissue; HeLa cells.
  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.21% BSA, 59% PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR10276(B)
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab167429 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 1/1000 - 1/10000. Predicted molecular weight: 110 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

ICC/IF 1/100 - 1/500.
IP 1/10 - 1/100.
Flow Cyt Use at an assay dependent concentration.

Target

  • Function
    Microtubule-dependent motor required for normal distribution of mitochondria and lysosomes.
  • Sequence similarities
    Belongs to the TRAFAC class myosin-kinesin ATPase superfamily. Kinesin family. Kinesin subfamily.
    Contains 1 kinesin motor domain.
  • Domain
    Composed of three structural domains: a large globular N-terminal domain which is responsible for the motor activity of kinesin (it hydrolyzes ATP and binds microtubule), a central alpha-helical coiled coil domain that mediates the heavy chain dimerization; and a small globular C-terminal domain which interacts with other proteins (such as the kinesin light chains), vesicles and membranous organelles.
  • Cellular localization
    Cytoplasm, cytoskeleton. Uniformly distributed between soma and neurites in hippocampal neurons.
  • Information by UniProt
  • Database links
  • Alternative names
    • Conventional kinesin heavy chain antibody
    • KIF 5B antibody
    • KIF5B antibody
    • Kinesin 1 antibody
    • kinesin 1 (110-120kD) antibody
    • Kinesin 1 heavy chain antibody
    • Kinesin family member 5B antibody
    • Kinesin heavy chain antibody
    • kinesin, heavy chain, ubiquitous antibody
    • Kinesin-1 heavy chain antibody
    • Kinesin1 antibody
    • KINH antibody
    • KINH_HUMAN antibody
    • KNS 1 antibody
    • KNS antibody
    • KNS1 antibody
    • Ubiquitous kinesin heavy chain antibody
    • UKHC antibody
    see all

Images

  • ab167429 (purified) at 1:20 dilution (2μg) immunoprecipitating KIF5B in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.

    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
    Lane 2 (+): ab167429 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab167429 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling KIF5B with purified ab167429 at 1:20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling KIF5B with Purified ab167429 at 1:250 dilution (0.07 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • All lanes : Anti-KIF5B antibody [EPR10276(B)] (ab167429) at 1/5000 dilution (purified)

    Lane 1 : Jurkat (human acute T cell leukemia) whole cell lysates
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell ) whole cell lysates
    Lane 3 : Mouse brain lysates
    Lane 4 : Rat brain lysates

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size : 110 kDa

    Blocking and diluting buffer: 5% NFDM/TBST



  • Predicted band size : 110 kDa

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: KIF5B knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab167429 observed at 110 kDa. Red - loading control, ab9484, observed at 37 kDa.

    Unpurified ab167429 was shown to specifically react with KIF5B in wild-type cells as signal was lost in KIF5B knockout cells. Wild-type and KIF5B knockout samples were subjected to SDS-PAGE. Ab167429 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/Immunofluorescence analysis HepG2(human hepatocellular carcinoma) labelling KIF5B with purified ab167429 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

  • All lanes : Anti-KIF5B antibody [EPR10276(B)] (ab167429) at 1/1000 dilution (unpurified)

    Lane 1 : Human fetal kidney lysate
    Lane 2 : HeLa lysate
    Lane 3 : Jurkat lysate
    Lane 4 : HepG2 lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size : 110 kDa
  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human brain tissue, labeling KIF5B using unpurified ab167429 at a 1/100 dilution.

  • Immunofluorescence analysis of HeLa cells, labeling KIF5B using unpurified ab167429 at a 1/100 dilution.

References

This product has been referenced in:
  • Stroud MJ  et al. Nesprin 1a2 is essential for mouse postnatal viability and nuclear positioning in skeletal muscle. J Cell Biol 216:1915-1924 (2017). Read more (PubMed: 28533284) »
  • Wang C  et al. Dynamic tubulation of mitochondria drives mitochondrial network formation. Cell Res 25:1108-20 (2015). Read more (PubMed: 26206315) »

See all 3 Publications for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (Adrenal carcinoma)
Specification
Adrenal carcinoma
Treatment
siRNA - 48 hours
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Verified customer

Submitted Nov 11 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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