Lipase A Antibody - BSA Free

Name: Lipase A Antibody - BSA Free
Brand: Novus Biologicals
SKU: NBP1-54155
Price: 429 USD
Availability: InStock
Rating: 5 (1 reviews)

Novus Biologicals | Catalog # NBP1-54155

(1)

Citations (8)

Product Documents

View Available Conjugates

100% Guarantee

Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

View all LIPA Primary Antibodies »

Product Specifications

Immunogen

Partial recombinant protein made to an internal region of the human Lipase A protein (within residues 150-300). [Swiss-Prot P38571]

Reactivity Notes

Immunogen sequence has 82% identity to pig and 73% identity to rat and cow.

Localization

Lysosome.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Lipase A Antibody - BSA Free

Western Blot: Lipase A AntibodyBSA Free [NBP1-54155]

Western Blot: Lipase A Antibody [NBP1-54155] - Analysis using NBP1-54155 on cellular extracts of human prostate adenoma tissue, DU145, PC3, and 22RV1 prostate cancer cell lines, and HFL1 Normal primary human fibroblasts. NBP1-54155 revealed a strong specific band at the expected molecular weight for Lipase A at 45 kDa in different lysates of human cancer cell lines. NBP1-54155 specifically reveals LIPA in extracts from human prostate adenoma tissue and prostate cancer cell lines DU145, PC3, and 22RV1.The protein bands were visible in the positive but not in the negative controls. Therefore, NBP1-54155 was found to be suitable for checking the expression levels of Lipase A protein. Image from verified customer review.

Immunocytochemistry/ Immunofluorescence: Lipase A Antibody - BSA Free [NBP1-54155]

Lipase-A-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-54155-img0008.jpg

Western Blot: Lipase A AntibodyBSA Free [NBP1-54155]

Lipase-A-Antibody-Western-Blot-NBP1-54155-img0009.jpg

Simple Western: Lipase A AntibodyBSA Free [NBP1-54155]

Simple Western: Lipase A Antibody [NBP1-54155] - Image shows a specific band for Lipase A in 0.5 mg/mL of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry/ Immunofluorescence: Lipase A Antibody - BSA Free [NBP1-54155]

Immunocytochemistry/Immunofluorescence: Lipase A Antibody [NBP1-54155] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Lipase A Antibody NBP1-54155 at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry-Paraffin: Lipase A Antibody - BSA Free [NBP1-54155]

Immunohistochemistry-Paraffin: Lipase A Antibody - BSA Free [NBP1-54155] - Analysis of a FFPE tissue section of human stomach using 1:200 dilution of Lipase A antibody (NBP1-54155). The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.

Western Blot: Lipase A AntibodyBSA Free [NBP1-54155]

Western Blot: Lipase A Antibody [NBP1-54155] - Detection of LIPA in HepG2 whole cell extract.

Immunohistochemistry: Lipase A Antibody - BSA Free [NBP1-54155]

Immunohistochemistry: Lipase A Antibody [NBP1-54155] - Staining of LIPA in mouse stomach smooth muscle.

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] - FoxO1-mediated lysosomal acid lipase (Lipa) induction in NR & Metf-treated 3T3-L1 adipocytes. (a) WB of FoxO1, ATGL & Lipa in total protein extracts from 3T3-L1 adipocytes at different times of NR. (b) RT-qPCR analysis of relative Lipa & ATGL mRNA levels in 3T3-L1 after 4 h from NR. Dashed line indicates mRNA value of controls. (c) After 4 h from NR, 3T3-L1 adipocytes refed w/ complete cell culture medium (CM) up to 8 h. Total protein extracts used for WB analysis of FoxO1 & Lipa. (d) WB of FoxO1 in total & nuclear protein extracts from 3T3-L1 adipocytes at different times of NR. (e) ChIP assay carried out on crosslinked nuclei from 3T3-L1 adipocytes subjected to NR for 4 h & Metf for 16 h by using FoxO1 antibody followed by qPCR analysis of FoxO1RE on Lipa promoter (−51 bp). Dashed line indicates IgG value. (f & g) 3T3-L1 adipocytes transfected w/ siRNA against FoxO1 (FoxO1(−)) or w/ a scramble siRNA (Scr). WB of FoxO1 & Lipa (f) & RT-qPCR analysis of relative Lipa mRNA level (g) performed in 3T3-L1 adipocytes 4 h after NR. (h) WB of FoxO1 & Lipa in 3T3-L1 adipocytes at different times of 5 mM Metformin (Metf) treatment. (i) Confocal analysis of FoxO1 localization in 3T3-L1 adipocytes treated w/ 5 mM Metf for 16 h. Nuclei stained w/ Hoechst 33342. Colocalization plugin (ImageJ Software) used to identify FoxO1-Hoechst colocalization (white spots). (j) RT-qPCR analysis of relative Lipa mRNA level performed in 3T3-L1 adipocytes treated w/ Metf for 16 h. (k) 3T3-L1 adipocytes transfected w/ siRNA against FoxO1 (FoxO1(−)) or w/ a scramble siRNA (Scr). WB of FoxO1 & Lipa performed in 3T3-L1 adipocytes treated w/ 5 mM Metf for 24 h. All values given as mean±S.D. (n=4). *P<0.05, **P<0.01 versus controls. °P<0.05 versus NR Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24136225), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] - AMPK drives Lipa-released FFAs oxidation restraining energetic catastrophe. (a) 3T3-L1 cells were transfected with DN-AMPK or empty vector. RT-qPCR analysis of relative peroxisome proliferator-activated receptor gamma-1 alpha, peroxisome proliferator-activated receptor-alpha, carnitine palmitoyltransferase 1b & acyl-CoA oxidase 1 mRNA levels were performed after 4 h of NR or 16 h of Metf treatment. Dashed line indicates the mRNA value of untreated DN-AMPK cells (Ctr). mRNA levels of untreated cells transfected with empty vector were similar to untreated DN-AMPK cells (data not shown). (b) Cheminoluminescent assay of ATP level in 3T3-L1 adipocytes transfected with DN-AMPK or empty vector after 8 h NR or 16 h Metf treatment. ATP level was expressed as pmol ATP per mg protein. (c) After 8 h of NR or 16 h Metf treatment, FFAs were enzymatically detected in culture medium of 3T3-L1 adipocytes transfected with DN-AMPK or empty vector. Values were expressed as μg FFAs per mg protein. (d) Left panel: western blot of AMPKpT172, PARP-1 & cleaved form of caspase-3 in 3T3-L1 adipocytes transfected with DN-AMPK or empty vector & subjected to 8 h NR. Right panel: cytofluorimetric analysis of apoptosis in DN-AMPK cells subjected to 8 h NR. (e) Western blot of PARP-1 & cleaved form of caspase-3 in 3T3-L1 adipocytes transfected with DN-AMPK or empty vector & treated with Metf for 16 h. (f) Western blot of FoxO1, Lipa, LC3 in 3T3-L1 adipocytes transfected with DN-AMPK or empty vector & subjected to 4 h NR. beta -actin was used as loading control. All values are given as mean±S.D. *P<0.05, **P<0.01 versus controls; °P<0.05, °°P<0.01 versus Metf treatment. All data are representative of at least three independent experiments Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24136225), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] - Lipa downregulation impairs lipid breakdown & elicits cell death in nutrient restricted adipocytes. (a) 3T3-L1 adipocytes were transfected with siRNA against Lipa (Lipa(−)) or with a scramble siRNA (Scr). Western blot of Lipa, PARP-1 & cleaved form of caspase-3 in total protein extracts from 3T3-L1 adipocytes after 4 h of NR. (b) TG content was quantified by ORO staining in fixed 3T3-L1 adipocytes 6 h after NR. (c) RT-qPCR analysis of relative peroxisome proliferator-activated receptor gamma-1 alpha, peroxisome proliferator-activated receptor-alpha & carnitine palmitoyltransferase 1b mRNA levels was performed in 3T3-L1 adipocytes 4 h after NR. (d) FFAs were analyzed in culture medium 6 h after NR. beta -actin was used as loading control. All values are given as mean±S.D. *P<0.05, **P<0.01 versus controls; °P<0.05 versus NR treatment. All data are representative of at least three independent experiments Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24136225), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Immunocytochemistry/ Immunofluorescence: Lipase A Antibody - BSA Free [NBP1-54155] -

Metabolic stress triggers lipophagy in adipocytes. (a) 3T3-L1 adipocytes were transfected with EGFP-LC3 expression vector (green) and subjected to NR or treated with Metf. Cells were immunostained with PLIN antibody (red). (b) 3T3-L1 adipocytes were subjected to NR or treated with Metf for 8 h. Cells were immunostained with Lipa (green) and PLIN (red) antibodies. (c) 3T3-L1 adipocytes were subjected to NR or treated with Metf for 8 h. Cells were immunostained with LAMP1 antibody (green). LDs were visualized by using Nile Red (red). Nuclei were counterstained with Hoechst 33342 (blue). Arrowheads indicate examples of colocalization points. Overlap coefficients were calculated by JACoP (plugin of ImageJ Software) in at least four different images. All values are given as mean+/-S.D. *P<0.05, **P<0.01 versus controls Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24136225), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

View 13 more images

Applications for Lipase A Antibody - BSA Free

Application

Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100 - 1:500

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Simple Western

1:10000

Western Blot

1 - 2 ug/ml

Application Notes

This LIPA antibody is useful for ICC/IF, IHC-P and Western blot, where a band is seen ~45 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HepG2 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:10,000, apparent MW was 75 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 1 review rated 5 using NBP1-54155 in the following applications:

Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: LIPA

Lipase A (LIPA) is an enzyme essential for biotransformation, sequestration and breakdown of cholesterol. It is localized in lysosomes and is expressed at high levels in small intestinal villi, the zona fasciculate/reticularis of the adrenal cortex, pancreatic acini, renal tubular epithelium, hepatocytes, spleenocytes and thymic cells. LIPA is highly crucial for hydrolysis of cholesteryl esters and triglycerides internalized via receptor-mediated endocytosis of lipoproteins. It is also implicated in mediating LDL uptake effects on suppression of HMG-CoA reductase and activation of endogenous cholesteryl ester synthesis. Defective LIPA in liver and kidney may lead to lipoprotein cholesterol accumulation which results in pathologies including atherosclerosis, inflammation, NIDDM and chronic kidney disease. Defects in LIPA are the cause of Wolman disease (WOD) and cholesteryl ester storage disease (CESD).

Long Name

Lipase A

Alternate Names

CESD, Lipase A

Entrez Gene IDs

3988 (Human); 16889 (Mouse)

Gene Symbol

LIPA

UniProt

P38571 (Human), Q9Z0M5 (Mouse)

Additional LIPA Products

Product Documents for Lipase A Antibody - BSA Free

Download Product Datasheets

Download ICC/IF Protocol

Download IHC-P Protocol

Download Western Blot Protocol

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for Lipase A Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Lipase A Antibody - BSA Free

Customer Reviews for Lipase A Antibody - BSA Free (1)

5 out of 5

1 Customer Rating

5 Stars

100%

4 Stars

3 Stars

2 Stars

1 Stars

Have you used Lipase A Antibody - BSA Free?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review

Customer Images

Showing 1 - 1 of 1 review Showing All

Filter By:

Lipase A Antibody

Name: Anonymous

Application: Western Blot

Sample Tested: prostate adenoma tissue and HFL1 primary human fibroblasts extract

Species: Human

Verified Customer | Posted 02/01/2018

Western blot analysis using NBP1-54155 on cellular extracts of human prostate adenoma tissue, DU145, PC3, and 22RV1 prostate cancer cell lines, and HFL1 Normal primary human fibroblasts.

Positive control: Human prostate adenoma tissue, DU145, PC3, and 22RV1 prostate cancer cell line lysates Negative control: HFL1 primary human fibroblasts extract 20µg/lane protein Incubate with primary rabbit anti-LIPA primary antibody (Novus Biologicals, NBP1-54155, lot A-4) diluted at 1:5000 in 3% TBS-T/BSA ON at 4°C. Incubate with secondary goat anti-rabbit/HRP-conjugated antibody (Bio-Rad, cat 1706515) diluted 1:3000 in 3% TBS-T/BSA for 1h-2h at RT. Image acquisition 30 sec exposure Strip membrane Incubate membrane with primary mouse beta-Actin loading control antibody (Novus Biologicals, NB600501, lot 64M4789V) diluted at 1:5000 in 3% TBS-T/BSA and with secondary anti-mouse/HRP-conjugated antibody (Bio-Rad, 170547) at 1:3000 in 3% TBS-T/BSA. Image acquisition 30 sec exposure Experimental notes & Conclusion: NBP1-54155 revealed a strong specific band at the expected molecular weight for Lipase A at 45kDa in different lysates of human cancer cell lines. The protein bands were visible in the positive but not in the negative controls. Therefore, NBP1-54155 was found to be suitable for checking the expression levels of Lipase A protein. Conclusion Passed. NBP1-54155 specifically reveals LIPA in extracts from human prostate adenoma tissue and prostate cancer cell lines DU145, PC3, and 22RV1. Validation lab name: Giannis Lamprou, Avgi Tsolou, Michael Koukourakis; Department of Radiation Oncology - Radiobiology - Radiopathology, Medical School - Democritus University of Thrace - University Hospital of Alexandroupolis Via antibodies-online Validation Program.

There are no reviews that match your criteria.

Showing 1 - 1 of 1 review Showing All

Protocols

View specific protocols for Lipase A Antibody - BSA Free (NBP1-54155):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

*Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs

No product specific FAQs exist for this product.

View all FAQs for Antibodies

Lipase A Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-54155

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

Format

Product Specifications

Immunogen

Reactivity Notes

Localization

Clonality

Host

Isotype

Scientific Data Images for Lipase A Antibody - BSA Free

Western Blot: Lipase A AntibodyBSA Free [NBP1-54155]

Immunocytochemistry/ Immunofluorescence: Lipase A Antibody - BSA Free [NBP1-54155]

Western Blot: Lipase A AntibodyBSA Free [NBP1-54155]

Simple Western: Lipase A AntibodyBSA Free [NBP1-54155]

Immunocytochemistry/ Immunofluorescence: Lipase A Antibody - BSA Free [NBP1-54155]

Immunohistochemistry-Paraffin: Lipase A Antibody - BSA Free [NBP1-54155]

Western Blot: Lipase A AntibodyBSA Free [NBP1-54155]

Immunohistochemistry: Lipase A Antibody - BSA Free [NBP1-54155]

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Western Blot: Lipase A Antibody - BSA Free [NBP1-54155] -

Immunocytochemistry/ Immunofluorescence: Lipase A Antibody - BSA Free [NBP1-54155] -

Applications for Lipase A Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence

Immunohistochemistry

Immunohistochemistry-Paraffin

Simple Western

Western Blot

Reviewed Applications

Formulation, Preparation, and Storage

Purification

Formulation

Format

Preservative

Concentration

Shipping

Stability & Storage

Background: LIPA

Long Name

Alternate Names

Entrez Gene IDs

Gene Symbol

UniProt

Additional LIPA Products

Product Documents for Lipase A Antibody - BSA Free

Product Datasheets

Protocols & Manuals

COA

Certificate of Analysis

SDS

Product Specific Notices for Lipase A Antibody - BSA Free

Citations for Lipase A Antibody - BSA Free

Customer Reviews for Lipase A Antibody - BSA Free (1)

Customer Images

Lipase A Antibody

Protocols

View specific protocols for Lipase A Antibody - BSA Free (NBP1-54155):

Immunocytochemistry/Immunofluorescence protocol for Lipase A Antibody (NBP1-54155)

Immunohistochemistry-Paraffin protocol for Lipase A Antibody (NBP1-54155)

Western Blot protocol specific for LIPA antibody (NBP1-

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs

Related Products