Overview

  • Product name

    Anti-DAGLA antibody
  • Description

    Goat polyclonal to DAGLA
  • Host species

    Goat
  • Tested applications

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

    Reacts with: Mouse, Rat, Human, Monkey
  • Immunogen

    Synthetic peptide:

    C-PAKQDELVISAR

    , corresponding to C terminal amino acids 1031-1042 of Human DAGLA.

  • Positive control

    • Human liver lysates (WB). Mouse cerebellum (IHC) This antibody gave a positive result when used in the following formaldehyde fixed cell lines: SKNSH.

Properties

Applications

Our Abpromise guarantee covers the use of ab81984 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
WB Use a concentration of 0.3 - 1 µg/ml. Predicted molecular weight: 115 kDa.
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 10 µg/ml.
IHC-P 1/500 - 1/5000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

Target

  • Function

    Catalyzes the hydrolysis of diacylglycerol (DAG) to 2-arachidonoyl-glycerol (2-AG), the most abundant endocannabinoid in tissues. Required for axonal growth during development and for retrograde synaptic signaling at mature synapses.
  • Tissue specificity

    Highly expressed in brain and pancreas.
  • Involvement in disease

    Defects in DAGLA may be a cause of spinocerebellar ataxia type 20 (SCA20) [MIM:608687]. Spinocerebellar ataxia is a clinically and genetically heterogeneous group of cerebellar disorders. Patients show progressive incoordination of gait and often poor coordination of hands, speech and eye movements, due to degeneration of the cerebellum with variable involvement of the brainstem and spinal cord. SCA20 is an autosomal dominant, adult-onset form characterized by dysarthria due to spasmodic dysphonia followed by slowly progressive ataxia. Note=A copy number variation consisting of a 260-kb duplication at chromosome 11q12.2-12.3 is responsible for SCA20. The critical gene within the duplicated segment may be DAGLA.
  • Sequence similarities

    Belongs to the AB hydrolase superfamily. Lipase family.
  • Cellular localization

    Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • C11orf11 antibody
    • Chromosome 11 open reading frame 11 antibody
    • DAGL(ALPHA) antibody
    • Dagla antibody
    • DAGLALPHA antibody
    • DGL alpha antibody
    • DGL-alpha antibody
    • DGLA_HUMAN antibody
    • Diacylglycerol lipase alpha antibody
    • KIAA0659 antibody
    • Neural stem cell derived dendrite regulator antibody
    • Neural stem cell-derived dendrite regulator antibody
    • NSDDR antibody
    • Sn1-specific diacylglycerol lipase alpha antibody
    see all

Images

  • Anti-DAGLA antibody (ab81984) at 0.3 µg/ml + human liver lysate (in RIPA buffer) at 35 µg

    Developed using the ECL technique.

    Predicted band size: 115 kDa
    Observed band size: 125 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 22 kDa, 55 kDa. We are unsure as to the identity of these extra bands.



    Some minor background is detected and is blocked by the immunizing peptide.
  • Immunohistochemistical detection of DAGLA using ab81984 on formaldehyde fixed rat cerebellum tissue sections (cerebellum). Antigen retrieval step: heat mediated in citric acid pH6. 1%BSA as blocking agent for 10 mins@ rt°C. Primary antibody dilution 1/1000 for 2 hours. Secondary antibody: anti-sheep IgG conjugated to biotin (1/200). The pattern of immunolabelling is very similar to that obtained in mouse. However, the cell bodies of some Purkinje cells are also positive, this appears to be punctate and evenly distributed.

    See Abreview

  • Immunohistochemistical detection of DAGLA with ab81984 on formaldehyde fixed mouse cerebellum sections. Antigen retrieval step: heat mediated in citric acid pH6. 1% BSA used as blocking agent for 10 mins @ rt°C. Primary antibody ab81984 incubated at 1/500 for 2 hours. Secondary antibody: anti sheep IgG conjugated to biotin (1/200). The upper image shows DAGL immunostaining with ab81984 on wildtype mouse cerebellum, the lower image shows an absence of DAGL alpha immunostaining on knockout mouse mouse cerebellum. These results are consistent with published data with other anti-DAGL alpha antibodies (http://jcb.rupress.org/cgi/content/full/163/3/463)

    See Abreview

  • ab81984 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal donkey 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 ab81984 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 donkey anti- goat 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 1.43µM.

References

This product has been referenced in:

  • Okubo Y  et al. Diacylglycerol lipase alpha promotes tumorigenesis in oral cancer by cell-cycle progression. Exp Cell Res 367:112-118 (2018). WB . Read more (PubMed: 29614312) »
  • Smaga I  et al. Changes in the Brain Endocannabinoid System in Rat Models of Depression. Neurotox Res 31:421-435 (2017). Read more (PubMed: 28247204) »
See all 3 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Immunoprecipitation
Immuno-precipitation step
Protein G
Sample
Mouse Tissue lysate - whole (brain)
Specification
brain
Total protein in input
10 µg

Ms. Ya Zhou

Verified customer

Submitted Dec 04 2013

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10)
Sample
Monkey Cell lysate - other (COS 7)
Specification
COS 7
Treatment
TRANSFECT DAGLA
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Sep 30 2013

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (HEK293)
Specification
HEK293
Permeabilization
Yes - 0.2% triton
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 26 2013

Application
Western blot
Sample
Mouse Tissue lysate - whole (olfactory epithelium)
Loading amount
40 µg
Specification
olfactory epithelium
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (12.5%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 05 2011

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Neural stem cell line)
Loading amount
30 µg
Specification
Neural stem cell line
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Apr 06 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Cerebellum)
Specification
Cerebellum
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C

Mr. Carl Hobbs

Verified customer

Submitted Jan 15 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (cerebellum)
Specification
cerebellum
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C

Mr. Carl Hobbs

Verified customer

Submitted Dec 15 2009

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Monkey Tissue sections (Marmoset cerebellum)
Specification
Marmoset cerebellum
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C

Mr. Carl Hobbs

Verified customer

Submitted Dec 15 2009

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