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    clasp1-antibody-epr3409-ab108620.pdf

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Cell Biology Cell Cycle Cell Division Spindle
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CLASP1 antibody [EPR3409] (ab108620)

  • Datasheet
Reviews (1) Submit a question References (7)

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Immunocytochemistry/ Immunofluorescence - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Immunocytochemistry/ Immunofluorescence - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Flow Cytometry - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Flow Cytometry - Anti-CLASP1 antibody [EPR3409] (ab108620)
  • Anti-CLASP1 antibody [EPR3409] (ab108620)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3409] to CLASP1
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

View more associated products

Overview

  • Product name

    Anti-CLASP1 antibody [EPR3409]
    See all CLASP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR3409] to CLASP1
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Mouse
    Human
    WB
    Rat
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: U87-MG, HeLa, T47-D and C6 cell lysates and rat brain tissue lysate. IHC-P: Human and mouse kidney and human cerebral cortex tissues. ICC/IF: HeLa cells. Flow Cyt: 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3409
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Spindle
    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Other cell division antibodies

Associated products

  • Alternative Versions

    • Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • T47D whole cell lysate (ab14899)
    • HeLa whole cell lysate (ab150035)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108620 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Mouse
Human
WB
Rat
Human
Application Abreviews Notes
ICC/IF
1/100.
WB (1)
1/5000 - 1/50000. Detects a band of approximately 160 kDa (predicted molecular weight: 169 kDa).
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
1/100.

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

Notes
ICC/IF
1/100.
WB
1/5000 - 1/50000. Detects a band of approximately 160 kDa (predicted molecular weight: 169 kDa).
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
1/100.

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

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Microtubule plus-end tracking protein that promotes the stabilization of dynamic microtubules. Involved in the nucleation of noncentrosomal microtubules originating from the trans-Golgi network (TGN). Required for the polarization of the cytoplasmic microtubule arrays in migrating cells towards the leading edge of the cell. May act at the cell cortex to enhance the frequency of rescue of depolymerizing microtubules by attaching their plus-ends to cortical platforms composed of ERC1 and PHLDB2. This cortical microtubule stabilizing activity is regulated at least in part by phosphatidylinositol 3-kinase signaling. Also performs a similar stabilizing function at the kinetochore which is essential for the bipolar alignment of chromosomes on the mitotic spindle.
    • Sequence similarities

      Belongs to the CLASP family.
      Contains 7 HEAT repeats.
    • Post-translational
      modifications

      Phosphorylated upon DNA damage, probably by ATM or ATR.
    • Cellular localization

      Cytoplasm > cytoskeleton. Cytoplasm > cytoskeleton > centrosome. Chromosome > centromere > kinetochore. Cytoplasm > cytoskeleton > spindle. Golgi apparatus > trans-Golgi network. Localizes to microtubule plus ends. Localizes to centrosomes, kinetochores and the mitotic spindle from prometaphase. Subsequently localizes to the spindle midzone from anaphase and to the midbody from telophase. In migrating cells localizes to the plus ends of microtubules within the cell body and to the entire microtubule lattice within the lamella. Localizes to the cell cortex and this requires ERC1 and PHLDB2.
    • Target information above from: UniProt accession Q7Z460 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 23332 Human
      • Entrez Gene: 76707 Mouse
      • Entrez Gene: 304740 Rat
      • Omim: 605852 Human
      • SwissProt: Q7Z460 Human
      • SwissProt: Q80TV8 Mouse
      • Unigene: 469840 Human
      • Unigene: 708183 Human
      • Unigene: 138740 Mouse
      see all
    • Alternative names

      • 1700030C23Rik antibody
      • 5730583A19Rik antibody
      • B130045P17Rik antibody
      • CLAP1_HUMAN antibody
      • clasp1 antibody
      • CLIP associating protein 1 antibody
      • CLIP associating protein CLASP1 antibody
      • CLIP-associating protein 1 antibody
      • Cytoplasmic linker associated protein 1 antibody
      • Cytoplasmic linker-associated protein 1 antibody
      • DKFZp686D1968 antibody
      • DKFZp686H2039 antibody
      • FLJ33821 antibody
      • FLJ41222 antibody
      • hOrbit1 antibody
      • KIAA0622 antibody
      • MAST1 antibody
      • MGC131895 antibody
      • mKIAA0622 antibody
      • Multiple asters 1 antibody
      • Multiple asters homolog 1 antibody
      • Protein Orbit homolog 1 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Immunocytochemistry/ Immunofluorescence - Anti-CLASP1 antibody [EPR3409] (ab108620)

      Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling CLASP1 (green) with ab108620 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/100) and secondary antibody, ab150120 Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    • Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
      All lanes : Anti-CLASP1 antibody [EPR3409] (ab108620) at 1/10000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : CLASP1 knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : Jurkat whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 169 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab108620 observed at 169 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab108620 was shown to recognize CLASP1 in wild-type HAP1 cells as signal was lost at the expected MW in CLASP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CLASP1 knockout samples were subjected to SDS-PAGE. Ab108620 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 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.

    • Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
      All lanes : Anti-CLASP1 antibody [EPR3409] (ab108620) at 1/20000 dilution (purified)

      Lane 1 : U87-MG cell lysate
      Lane 2 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 169 kDa
      Observed band size: 160 kDa
      why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Anti-CLASP1 antibody [EPR3409] (ab108620) at 1/50000 dilution (purified) + Rat brain tissue lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 169 kDa
      Observed band size: 160 kDa why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Western blot - Anti-CLASP1 antibody [EPR3409] (ab108620)
      All lanes : Anti-CLASP1 antibody [EPR3409] (ab108620) at 1/10000 dilution (unpurified)

      Lane 1 : T47-D cell lysate
      Lane 2 : C6 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 169 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue labelling CLASP1 with purified ab108620 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labeling CLASP1 with purified ab108620 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] (ab108620)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling CLASP1 with unpurified ab108620 at 1/250.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Immunocytochemistry/ Immunofluorescence - Anti-CLASP1 antibody [EPR3409] (ab108620)

      Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling CLASP1 with unpurified ab108620 at 1/100.

    • Flow Cytometry - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Flow Cytometry - Anti-CLASP1 antibody [EPR3409] (ab108620)

      Flow cytometry analysis of HeLa cells labelling CLASP1 with purified ab108620 at 1/50 (red). Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • Flow Cytometry - Anti-CLASP1 antibody [EPR3409] (ab108620)
      Flow Cytometry - Anti-CLASP1 antibody [EPR3409] (ab108620)

      Overlay histogram showing HeLa cells stained with unpurified ab108620 (red line). The cells were fixed with 80% methanol (5 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 (ab108620, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    • Anti-CLASP1 antibody [EPR3409] (ab108620)
      Anti-CLASP1 antibody [EPR3409] (ab108620)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (7)

    Publishing research using ab108620? Please let us know so that we can cite the reference in this datasheet.

    ab108620 has been referenced in 7 publications.

    • Sonntag T  et al. The KLDpT activation loop motif is critical for MARK kinase activity. PLoS One 14:e0225727 (2019). PubMed: 31794565
    • Procter DJ  et al. The HCMV Assembly Compartment Is a Dynamic Golgi-Derived MTOC that Controls Nuclear Rotation and Virus Spread. Dev Cell 45:83-100.e7 (2018). PubMed: 29634939
    • Qin Z  et al. MicroRNAs Provide Feedback Regulation of Epithelial-Mesenchymal Transition Induced by Growth Factors. J Cell Physiol 231:120-9 (2016). PubMed: 26032086
    • Amin MA  et al. CLIP-170 recruits PLK1 to kinetochores during early mitosis for chromosome alignment. J Cell Sci 127:2818-24 (2014). ICC/IF ; Human . PubMed: 24777477
    • Zhao X  et al. Host microtubule plus-end binding protein CLASP1 influences sequential steps in the Trypanosoma cruzi infection process. Cell Microbiol 15:571-84 (2013). PubMed: 23107073
    • Naghavi MH  et al. Plus-end tracking proteins, CLASPs, and a viral Akt mimic regulate herpesvirus-induced stable microtubule formation and virus spread. Proc Natl Acad Sci U S A 110:18268-73 (2013). PubMed: 24145430
    • Chen X  et al. Analysis of substrates of protein kinase C isoforms in human breast cells by the traceable kinase method. Biochemistry 51:7087-97 (2012). PubMed: 22897107

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Western blot abreview for Anti-CLASP1 antibody [EPR3409]

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Specification
    HEK cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Ms. Jessie Bertram

    Verified customer

    Submitted Aug 15 2019

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