Recombinant 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: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-CLASP1 antibody [EPR3409]
See all CLASP1 primary antibodies -
Description
Rabbit monoclonal [EPR3409] to CLASP1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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 (intra): HeLa cells.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3409 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/100.
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WB | (1) |
1/5000 - 1/50000. Detects a band of approximately 160 kDa (predicted molecular weight: 169 kDa).
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IHC-P |
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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. |
Target
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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
modificationsPhosphorylated 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. - Information by UniProt
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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
see all -
Alternative names
- 1700030C23Rik antibody
- 5730583A19Rik antibody
- B130045P17Rik antibody
see all
Images
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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).
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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 kDaLanes 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. -
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.
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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.
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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) 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.
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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.
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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.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling CLASP1 with unpurified ab108620 at 1/100.
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Intracellular 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.
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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 Fluorr® 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (11)
ab108620 has been referenced in 11 publications.
- Lau EO et al. DIAPH3 deficiency links microtubules to mitotic errors, defective neurogenesis, and brain dysfunction. Elife 10:N/A (2021). PubMed: 33899739
- González-Barriga A et al. Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1. Front Cell Neurosci 15:662035 (2021). PubMed: 34025359
- Tsuchiya K & Goshima G Microtubule-associated proteins promote microtubule generation in the absence of γ-tubulin in human colon cancer cells. J Cell Biol 220:N/A (2021). PubMed: 34779859
- Maurin J et al. The Beta-Tubulin Isotype TUBB6 Controls Microtubule and Actin Dynamics in Osteoclasts. Front Cell Dev Biol 9:778887 (2021). PubMed: 34869381
- Sonntag T et al. The KLDpT activation loop motif is critical for MARK kinase activity. PLoS One 14:e0225727 (2019). PubMed: 31794565