Overview

  • Product name
  • Description
    Rabbit polyclonal to DCAMKL1
  • Specificity
    This antibody recognizes 4 different human isoforms (AS: 81.1 KDa, AL: 82.2 KDa, BS: 46.5 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 conjugated to KLH derived from within residues 700 to the C-terminus of Mouse DCAMLK1.

    (Peptide available as ab31703.)

  • Positive control
    • WB: Mouse Brain, Rat Brain, Human Brain IHC: Rat brain (e.g. cortex, striatum)

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

Anti-DCAMKL1 antibody images



  • Predicted band size : 47, 82 kDa

    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 cells as signal was lost in DCAMKL1 knockout cells. 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 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • ICC/IF image of ab31704 stained PC12 cells. The cells were 100% methanol fixed (5 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, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas 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) at a concentration of 1.43æM.

  • 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

  • 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
    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
  • 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
    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 minutesThe 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 for Anti-DCAMKL1 antibody (ab31704)

This product has been referenced in:
  • Brenna Ø  et al. Cellular localization of guanylin and uroguanylin mRNAs in human and rat duodenal and colonic mucosa. Cell Tissue Res 365:331-41 (2016). IHC-P ; Human . Read more (PubMed: 27044258) »
  • von Moltke J  et al. Tuft-cell-derived IL-25 regulates an intestinal ILC2-epithelial response circuit. Nature 529:221-5 (2016). Read more (PubMed: 26675736) »

See all 17 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Aug 02 2013

Thank you for your phone call.

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

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


Please find herethe discount code and more detailed informat...

Read More

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.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain sections)
Specification
Brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Username

Dr. Sophie Pezet

Verified customer

Submitted Dec 03 2009

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

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