Overview

  • Product name
    Anti-Lysosomal acid lipase antibody [9G7F12]
    See all Lysosomal acid lipase primary antibodies
  • Description
    Mouse monoclonal [9G7F12] to Lysosomal acid lipase
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, WB, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Ni-NTA purified truncated recombinant LAL expressed in E. Coli strain BL21 (DE3).

Properties

Applications

Our Abpromise guarantee covers the use of ab36597 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
Flow Cyt Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

WB 1/500 - 1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 46 kDa).
ELISA 1/10000.
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function
    Crucial for the intracellular hydrolysis of cholesteryl esters and triglycerides that have been internalized via receptor-mediated endocytosis of lipoprotein particles. Important in mediating the effect of LDL (low density lipoprotein) uptake on suppression of hydroxymethylglutaryl-CoA reductase and activation of endogenous cellular cholesteryl ester formation.
  • Involvement in disease
    Defects in LIPA are the cause of Wolman disease (WOD) [MIM:278000]. WOD is a severe manifestation of LIPA deficiency, leading to the accumulation of cholesteryl esters and triglycerides in most tissues of the body. WOD occurs in infancy and is nearly always fatal before the age of 1 year.
    Defects in LIPA are the cause of cholesteryl ester storage disease (CESD) [MIM:278000]. CESD is a mild manifestation of LIPA deficiency, leading to the accumulation of cholesteryl esters and triglycerides in most tissues of the body. It is characterized by late-onset.
  • Sequence similarities
    Belongs to the AB hydrolase superfamily. Lipase family.
  • Cellular localization
    Lysosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • Acid cholesteryl ester hydrolase antibody
    • CESD antibody
    • cholesterol ester hydrolase antibody
    • cholesterol ester storage disease antibody
    • Cholesteryl esterase antibody
    • Hydrolase deficiency antibody
    • LAL antibody
    • LAL deficiency cholesterol ester antibody
    • LICH_HUMAN antibody
    • lipA antibody
    • LIPA deficiency antibody
    • Lipase A antibody
    • lipase A, lysosomal acid, cholesterol esterase antibody
    • lysosomal acid lipase antibody
    • lysosomal acid lipase deficiency antibody
    • Lysosomal acid lipase/cholesteryl ester hydrolase antibody
    • Sterol esterase antibody
    see all

Images

  • All lanes : Anti-Lysosomal acid lipase antibody [9G7F12] (ab36597)

    Lane 1 : Molecular weight marker
    Lane 2 : Truncated LAL recombinant protein

    Predicted band size: 46 kDa
    Observed band size: ~45 kDa
    why is the actual band size different from the predicted?

  • ab36597 (2 µg/ml) staining lysosomal acid lipase in human lung using an automated system (DAKO Autostainer Plus). Using this protocol there is lysosomal staining in the bronchiolar epithelium and resident macrophages .
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Overlay histogram showing HepG2 cells stained with ab36597 (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 (ab36597, 0.5µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • Wang F  et al. Role of lysosomal acid lipase in the intracellular metabolism of LDL-transported dehydroepiandrosterone-fatty acyl esters. Am J Physiol Endocrinol Metab 295:E1455-61 (2008). WB ; Human . Read more (PubMed: 18796546) »
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for contacting us.

We do have LIPA protein available in catalogue. The ID is ab152501; https://www.abcam.com/lysosomal-acid-lipase-protein-tagged-ab152501.html

I have checked the image you have kindly provided, there is indeed bands at 45kDa and ˜65 kDa in HepG2 lane. Please also be aware that there are two isoforms of this protein, one has predicted molecular weight 39kDa and other 45kDa and due to heavy glycosylation the observed band size will be higher than 39 and 45. In HepG2 cells line the two bands at ˜45kDa and ˜65 kDa seems correspond to two isoforms, (http://www.uniprot.org/uniprot/P38571) so further tests about the identity of these isoform are recommended.

Please also cross check the negative LIPA protein expression in publications in mouse cells.

Though you have used the same antibody in past however we should expect some lot to lot variation so we always recommend our customers trying different dilutions of primary or secondary antibodies. The other important step is lysis of the cells, changing lysis buffer(ice cold) sometime helps. Pleas elaos make sure the lysis buffer contains protease inhibitors.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

This antibody has been tested with HepG2 cells and lung tissue sections; these samples will form a good positive control. Please find attached links to Human Protein Atlas; the database shows different tissue sections and cell lines in which this protein is highly expressed.

http://www.proteinatlas.org/ENSG00000107798

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Application
Western blot
Sample
Mouse Cell lysate - whole cell (J774 mouse macrophages, Human fibroblasts)
Loading amount
50000 cells
Specification
J774 mouse macrophages, Human fibroblasts
Treatment
lysis with LDS sample buffer, DTT, 70C for 10 min.
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

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Verified customer

Submitted Jun 15 2007

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