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DCAMKL1 Recombinant Rabbit Monoclonal Antibody [JA11-03] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750403

DCAMKL1 Recombinant Rabbit Monoclonal Antibody [JA11-03] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

DCAMKL1 Recombinant Rabbit Monoclonal Antibody [JA11-03] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human DCAMKL1 aa 649-698 / 740.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 82 kDa

Positive Control

SH-SY5Y cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, SH-SY5Y, rat brain tissue.

Conjugation

unconjugated

Clone Number

JA11-03

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

Lissencephaly (smooth brain) is an abnormality of brain development characterized by incomplete neuronal migration and a smooth cerebral surface, manifesting as severe mental retardation. Genetic analysis has identified two proteins that are mutated in some cases of lissencephaly, designated lissencephaly-1 protein (LIS1) and doublecortin. LIS1 displays sequence homology to β-subunits of heterotrimeric G proteins, and doublecortin contains a consensus Abl phosphorylation site. In addition, the DCAMKL1 (doublecortinlike and CAM kinase-like 1) protein shows homology to doublecortin. All three proteins are highly expressed in developing brain and may function together to regulate microtubules involved in neuronal migration. The DCAMKL1 protein encodes a functional kinase that is capable of phosphorylating myelin basic protein and itself, but its kinase activity does not appear to affect its microtubule polymerization activity.

Background References

1. Qu D et al. Doublecortin-like kinase 1 is elevated serologically in pancreatic ductal adenocarcinoma and widely expressed on circulating tumor cells. PLoS One 10:e0118933 (2015).

2. O\'Connell MR et al. Epigenetic changes and alternate promoter usage by human colon cancers for expressing DCLK1-isoforms: Clinical Implications. Sci Rep 5:14983 (2015).

Sequence Similarity

Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.

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.

Subcellular Location

Plasma membrane.

UNIPROT

SwissProt: O15075 Human

SwissProt: Q9JLM8 Mouse

SwissProt: O08875 Rat

Synonyms

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

Expand

Images

  • Western blot analysis of DCAMKL1 on different lysates with Rabbit anti-DCAMKL1 antibody (<a href="/products/HA750403" style="font-weight: bold;text-decoration: underline;">HA750403</a>) at 1/1,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 2: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 3: C6 cell lysate (20 µg/Lane)<br />Lane 4: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 5: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 82 kDa<br />Observed band size: 82/47 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750403" style="font-weight: bold;text-decoration: underline;">HA750403</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of DCAMKL1 on different lysates with Rabbit anti-DCAMKL1 antibody (HA750403) at 1/1,000 dilution.

    Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 3: C6 cell lysate (20 µg/Lane)
    Lane 4: Mouse brain tissue lysate (40 µg/Lane)
    Lane 5: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 82 kDa
    Observed band size: 82/47 kDa

    Exposure time: 6 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750403) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of SH-SY5Y cells labeling DCAMKL1 with Rabbit anti-DCAMKL1 antibody (<a href="/products/HA750403" style="font-weight: bold;text-decoration: underline;">HA750403</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DCAMKL1 antibody (<a href="/products/HA750403" style="font-weight: bold;text-decoration: underline;">HA750403</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of SH-SY5Y cells labeling DCAMKL1 with Rabbit anti-DCAMKL1 antibody (HA750403) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DCAMKL1 antibody (HA750403) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-DCAMKL1 antibody (<a href="/products/HA750403" style="font-weight: bold;text-decoration: underline;">HA750403</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750403" style="font-weight: bold;text-decoration: underline;">HA750403</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-DCAMKL1 antibody (HA750403) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750403) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of SH-SY5Y cells labeling DCAMKL1.<br /><br />Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (<a href="/products/HA750403" style="font-weight: bold;text-decoration: underline;">HA750403</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of SH-SY5Y cells labeling DCAMKL1.

    Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA750403, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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