Overview

  • Product name

  • Description

    Rabbit polyclonal to DCAMKL1
  • Host species

    Rabbit
  • Specificity

    This antibody recognizes 2 different human isoforms (AL: 82.2 KDa and BL: 47.6 KDa)
  • Tested applications

    Suitable for: ICC/IF, IHC-FoFr, IHC-P, WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse DCAMKL1 aa 700 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab31703)

  • Positive control

    • WB: Mouse Brain, Rat Brain, Human Brain IHC: Rat brain (e.g. cortex, striatum) ICC/IF: PC12 cell line

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab31704 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
ICC/IF Use a concentration of 1 µg/ml.
IHC-FoFr 1/1000.
IHC-P Use at an assay dependent concentration. PubMed: 20522640
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 47, 82 kDa (predicted molecular weight: 47, 82 kDa).Can be blocked with Mouse DCAMKL1 peptide (ab31703).
IP Use at an assay dependent concentration.

Target

  • Function

    Probable kinase that may be involved in a calcium-signaling pathway controlling neuronal migration in the developing brain. May also participate in functions of the mature nervous system.
  • Tissue specificity

    In fetal tissues, highly expressed in brain, detectable in lung and liver, but not in kidney. In adult tissues, expressed ubiquitously in the brain, detectable in the heart, liver, spleen, thymus, prostate, testis, ovary, small intestine and colon. The type A isoforms seem to be expressed predominantly in fetal brain whereas type B isoforms are expressed abundantly in both fetal and adult brain.
  • Sequence similarities

    Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.
    Contains 2 doublecortin domains.
    Contains 1 protein kinase domain.
  • Information by UniProt
  • Database links

  • Alternative names

    • Calcium/calmodulin-dependent protein kinase type I-like CPG16 antibody
    • CL1 antibody
    • CLICK1 antibody
    • Cpg16 antibody
    • DCDC3A antibody
    • Dcl antibody
    • Dclk antibody
    • Dclk1 antibody
    • DCLK1_HUMAN antibody
    • Doublecortin domain-containing protein 3A antibody
    • Doublecortin-like and CAM kinase-like 1 antibody
    • Doublecortin-like kinase 1 antibody
    • KIAA0369 antibody
    • Serine/threonine-protein kinase DCAMKL1 antibody
    • Serine/threonine-protein kinase DCLK1 antibody
    see all

Images

  • ab31704 at a 1/1000 dilution staining DCAMKL1 in rat brain tissue sections (hippocampus) by Immunohistochemistry/ PFA perfused, frozen sections, incubated for 18 hours at 20°C in PBS + 0.3% Triton X-100. Secondary used at 1/1000 polyclonal Goat anti-rabbit conjugated to Alexa Fluor 488.
    The immunostaining was performed using the `free floating` technique, using direct fluorescence. The antibody beautifully stains tracts of fibers in many brain areas such as the cortex, striatum. The image shown is staining observed at the level of the striatum.

    See Abreview

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: DCAMKL1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: NIH3T3 whole cell lysate (20 µg)
    Lane 4: Human brain whole tissue lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab31704 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab31704 was shown to specifically react with DCAMKL1 in wild-type HAP1 cells along with additional cross reactive bands. No bands were observed when DCAMKL1 knockout cells were examined. Wild-type and DCAMKL1 knockout samples were subjected to SDS-PAGE. ab31704 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • ab31704 stained in PC12 cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab31704 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • DCAMKL1 was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to DCAMKL1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31704.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 82kDa: DCAMKL1; non specific - 52 and 27kDa: We are unsure as to the identity of this extra band.
  • Anti-DCAMKL1 antibody (ab31704) at 1 µg/ml + Mouse Brain Whole Tissue Lysate at 20 µg

    Secondary
    IR Dye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

    Performed under reducing conditions.

    Predicted band size: 47, 82 kDa
    Observed band size: 47,82 kDa
    why is the actual band size different from the predicted?



    The 82 kDa and 47 kDa bands correspond to the AL and BL isoforms respectively. This antibody should not detect the AS and BS isoforms of DCAMLK1.
  • ICC/IF image of ab31704 stained PC12 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab31704, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
  • Anti-DCAMKL1 antibody (ab31704) at 1 µg/ml + Brain (Rat) Tissue Lysate - normal tissue at 10 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 47, 82 kDa
    Observed band size: 47,82 kDa why is the actual band size different from the predicted?

  • Anti-DCAMKL1 antibody (ab31704) at 1 µg/ml + Human brain tissue lysate - total protein (ab29466) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 47, 82 kDa
    Observed band size: 47,82 kDa why is the actual band size different from the predicted?
    Additional bands at: 30 kDa, 52 kDa (possible post-translational modification), 54 kDa (possible post-translational modification). We are unsure as to the identity of these extra bands.


    Exposure time: 5 minutes


    The 82 kDa and 47 kDa bands correspond to the AL and BL isoforms respectively. DCAMLK1 has a number of potential phosphorylation sites which may explain the higher migrating bands at 52 and 54 kDa.

References

This product has been referenced in:

  • Roy BC  et al. Co-localization of autophagy-related protein p62 with cancer stem cell marker dclk1 may hamper dclk1's elimination during colon cancer development and progression. Oncotarget 10:2340-2354 (2019). Read more (PubMed: 31040926) »
  • Liu R  et al. Constitutive STAT5 activation regulates Paneth and Paneth-like cells to control Clostridium difficile colitis. Life Sci Alliance 2:N/A (2019). Read more (PubMed: 30948494) »
See all 37 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Bone marrow cells)
Permeabilization
No
Specification
Bone marrow cells
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Mar 13 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric buffer
Permeabilization
No
Specification
lung
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Formaldehyde

Saltanat Ualiyeva

Verified customer

Submitted Dec 11 2018

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (12% SDS-PAGE)
Sample
Mouse Tissue lysate - other (Brain, Cortex)
Specification
Brain, Cortex
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 02 2013

Question
Answer

Thank you for your phone call.

A suitable isotype control for ab31704 and ab37994 would be: ab27478

https://www.abcam.com/Rabbit-polyclonal-IgG-ab27478.html


Please find herethe discount code and more detailed information for trying ab31704 in flow cytometry:

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab31704 in flow cytometry. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for flow cytometryand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

There are two monoclonals which are specific for the AS isoform: 1. ab88484- The immunogen has 52% homology with AL isoform. 2. ab56482- has the same immunogen but it has only been tested against the immunogen in WB.

Read More
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain sections)
Specification
Brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No

Dr. Sophie Pezet

Verified customer

Submitted Dec 03 2009

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