Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-KLC1 antibody [EPR12441(B)] (ab174273)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12441(B)] to KLC1
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-KLC1 antibody [EPR12441(B)]
    See all KLC1 primary antibodies

  • Description

    Rabbit monoclonal [EPR12441(B)] to KLC1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human KLC1 aa 500 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
    Database link: Q07866

  • Positive control

    • U87-MG, 293T, SH-SY5Y, and HeLa whole cell lysate (ab150035); Human brain tissue; U87-MG and 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab174273 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. Detects a band of approximately 61-70 kDa (predicted molecular weight: 65 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/50 - 1/500.
IP 1/10 - 1/100.
Flow Cyt 1/10 - 1/100.

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

 

Target

  • Function

    Kinesin is a microtubule-associated force-producing protein that may play a role in organelle transport. The light chain may function in coupling of cargo to the heavy chain or in the modulation of its ATPase activity.

  • Tissue specificity

    Found in a variety of tissues. Mostly abundant in brain and spine.

  • Sequence similarities

    Belongs to the kinesin light chain family.
    Contains 6 TPR repeats.

  • Post-translational
    modifications

    Isoform I is phosphorylated on Ser-600. Isoform J is phosphorylated on Ser-631.

  • Cellular localization

    Cytoplasm > cytoskeleton.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Kinesin 2 60/70kDa antibody
    • Kinesin light chain 1 antibody
    • Kinesin2 antibody
    • KLC 1 antibody
    • KLC antibody
    • Klc1 antibody
    • KLC1_HUMAN antibody
    • KNS 2 antibody
    • KNS 2A antibody
    • KNS2 antibody
    • KNS2A antibody
    • Medulloblastoma antigen MU MB 2.50 antibody
    • MGC15245 antibody
    see all

Images

  • Western blot - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    Western blot - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    All lanes : Anti-KLC1 antibody [EPR12441(B)] (ab174273) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : KLC1 knockout HeLa cell lysate
    Lane 3 : U-87 MG cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 65 kDa
    Observed band size: 65 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab174273 observed at 65 kDa. Red - loading control ab8245 observed at 36 kDa.

    ab174273 Anti-KLC1 antibody [EPR12441(B)] was shown to specifically react with KLC1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265284 (knockout cell lysate ab257496) was used. Wild-type and KLC1 knockout samples were subjected to SDS-PAGE. ab174273 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.

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    Immunocytochemistry/ Immunofluorescence - Anti-KLC1 antibody [EPR12441(B)] (ab174273)

    Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling KLC1 with purified ab174273 at 1/500. Cells were fixed with 100% methanol. 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLC1 antibody [EPR12441(B)] (ab174273)

    Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling KLC1 with ab174273 at 1/50 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    Western blot - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    All lanes : Anti-KLC1 antibody [EPR12441(B)] (ab174273) at 1/1000 dilution

    Lane 1 : U87-MG cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : SH-SY5Y cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 65 kDa

  • Immunoprecipitation - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    Immunoprecipitation - Anti-KLC1 antibody [EPR12441(B)] (ab174273)

    Western blot analysis on immunoprecipitation pellet from SH-SY5Y cell lysate using ab174273 at 1/10 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    Immunocytochemistry/ Immunofluorescence - Anti-KLC1 antibody [EPR12441(B)] (ab174273)

    Immunofluorescent analysis of U87-MG cells labeling KLC1 with ab174273 at 1/50 dilution.

  • Flow Cytometry - Anti-KLC1 antibody [EPR12441(B)] (ab174273)
    Flow Cytometry - Anti-KLC1 antibody [EPR12441(B)] (ab174273)

    Flow Cytometric analysis of permeabilized HeLa cells labeling KLC1 with ab174273 at 1/10 dilution (red) or a rabbit IgG (negative) (green).

References (4)

Publishing research using ab174273? Please let us know so that we can cite the reference in this datasheet.

ab174273 has been referenced in 4 publications.

  • Franchini DM  et al. Microtubule-Driven Stress Granule Dynamics Regulate Inhibitory Immune Checkpoint Expression in T Cells. Cell Rep 26:94-107.e7 (2019). PubMed: 30605689
  • Moamer A  et al. A role for kinesin-1 subunits KIF5B/KLC1 in regulating epithelial mesenchymal plasticity in breast tumorigenesis. EBioMedicine 45:92-107 (2019). PubMed: 31204277
  • Malikov V & Naghavi MH Localized Phosphorylation of a Kinesin-1 Adaptor by a Capsid-Associated Kinase Regulates HIV-1 Motility and Uncoating. Cell Rep 20:2792-2799 (2017). PubMed: 28930676
  • Wilson MH & Holzbaur EL Nesprins anchor kinesin-1 motors to the nucleus to drive nuclear distribution in muscle cells. Development 142:218-28 (2015). PubMed: 25516977

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Immunohistochemistry (Frozen sections) abreview for Anti-KLC1 antibody [EPR12441(B)]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Heart)
Permeabilization
Yes - Tween-20
Specification
Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Submitted Aug 12 2020

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