Perilipin-2/ADFP Antibody - BSA Free
Novus Biologicals | Catalog # NB110-40877
Key Product Details
Validated by
Orthogonal Validation, Biological Validation
Species Reactivity
Validated:
Human, Mouse, Rat, Bovine
Cited:
Human, Mouse, Rat, Bovine, Plant
Applications
Validated:
Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Dual RNAscope ISH-IHC, Immunocytochemistry/ Immunofluorescence, Simple Western
Cited:
Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC, SDS-Page
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Format
BSA Free
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Product Specifications
Immunogen
A synthetic peptide made to a C-terminal region of mouse ADFP (within residues 350-425) [Swiss-Prot# P43883]
Reactivity Notes
Rat reactivity reported in scientific literature (PMID: 29053516). Use in Bovine reported in scientific literature (PMID:32647143).
Localization
Membrane
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Theoretical MW
51 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for Perilipin-2/ADFP Antibody - BSA Free
Dual RNAscope ISH-IHC Analysis of Perilipin-2/ADFP in Mouse Intestine
Formalin-fixed paraffin-embedded tissue sections of mouse intestine were probed for Perilipin-2/ADFP mRNA (ACD RNAScope Probe, catalog #577111; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB110-40877) at 1:150 dilution with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to intestinal villi.
Simple Western Analysis of Perilipin-2/ADFP in HepG2 Cell Lysate
Simple Western lane view shows a specific band for ADFP in 0.5 mg/ml of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Immunohistochemical Staining of Perilipin-2/ADFP in Mouse Liver Tissue
ADFP antibody was tested in mouse liver using DAB with hematoxylin counterstain.
Immunocytochemistry/Immunofluorescence Staining of Perilipin-2/ADFP in NIH3T3 Cells
NIH3T3 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-Perilipin-2/ADFP Antibody NB110-40877 at 1 ug/ml overnight at 4C and detected with an anti-rabbit 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.
Western Blot Detection of Perilipin-2/ADFP in Mouse Liver Lysate
Detection of murine ADFP in mouse liver lysate.
Immunocytochemistry/Immunofluorescence Staining of Perilipin-2/ADFP in HepG2 Cells
HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-Perilipin-2/ADFP at a 1:100 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunocytochemistry/Immunofluorescence Analysis of Perilipin-2/ADFP in HepG2 Cells
HepG2 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-Perilipin-2/ADFP Antibody NB110-40877 at 1 ug/ml overnight at 4C and detected with an anti-rabbit 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.
Western Blot: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] -
Western Blot: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] - Western blot images (A) & quantifications (B–I) of muscle samples from WT & UCHL1 smKO mice for lipase ATGL (B), HSL (C), & MAGL (D), the fatty acid transport protein CD36 (E), the key lipid synthesis enzyme DGAT2 (F), perilipin 3 (G), perilipin 2 (H), & UCHL1 (I). n = 4-5 per group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35464088), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] -
Western Blot: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] - Western blot images (A) & quantifications (B–G) of cell lysates from C2C12 myotubes with control or UCHL1 siRNA knockdown for perilipin 2 (B), perilipin 5 (C), CD36 (D), HSL (E), MAGL (F), & SDHB (G). n = 4 per group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35464088), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Immunocytochemistry/ Immunofluorescence: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] -
Immunocytochemistry/ Immunofluorescence: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] - Lack of MT4-MMP in patrolling monocytes leads to the accumulation of Mafb+AIM+ macrophages in incipient atherosclerotic plaques. a Representative images of transverse sections of aortic sinus from Ldlr–/– mice transplanted with MT4-MMP+/+ (MT4+/+) or MT4-MMP–/– (MT4–/–) BM cells & fed a HFD for 1 week; sections were labeled for Mac3 (green), Mafb (red), & AIM (white), & with Hoechst (blue; nuclei); scale bar, 10 µm. b Number of Mac3+ cells (left), Mac3+Mafb+ cells (middle), & Mac3+Mafb+AIM+ cells (right) in the plaques of BM-transplanted Ldlr–/– mice fed a HFD for 1 week. c Representative images of transverse sections of aortic sinus from BM-transplanted Ldlr–/– mice fed a HFD for 1 week; sections were labeled for Mac3 (green), Mafb (red), & adipophilin (white), & with Hoechst (blue; nuclei); scale bar, 10 µm. d Relative % of adipophilin-positive & adipophilin-negative cells within the Mac3+Mafb+ population of BM-transplanted Ldlr–/– mice (1 week on HFD); n = 7 mice per genotype in two independent experiments. Data were tested by two-tailed Student’s t-test in b & by Fisher’s exact test in d. Results are expressed as mean ± SEM. *p < 0.05 Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/s41467-018-03351-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Perililpin-2/ADFP in HepG2 Human Cell Line.
Perililpin-2/ADFP was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rabbit anti-Perililpin-2/ADFP Affinity Purified Polyclonal Antibody conjugated to Biotin (Catalog # NB110-40877B) at 5 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.Applications for Perilipin-2/ADFP Antibody - BSA Free
Application
Recommended Usage
Immunocytochemistry/ Immunofluorescence
1:50-1:200
Immunohistochemistry-Frozen
reported in scientific literature (PMID 35041644)
Immunohistochemistry-Paraffin
1:200
Simple Western
1:50
Western Blot
2 ug/ml
Application Notes
This ADFP antibody can be used for Western blot and Immunofluorescence/Immunocytochemistry. In Western blot a band is observed at approx. 51 kDa. In ICC/IF, membrane staining was observed in HeLa cells. In IHC-P, staining was observed on the membrane of mouse liver cells 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 1:50, apparent MW was 77 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
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 1:50, apparent MW was 77 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
Reviewed Applications
Read 1 review rated 5 using NB110-40877 in the following applications:
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
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Background: Perilipin-2
References
1. Itabe H, Yamaguchi T, Nimura S, Sasabe N. Perilipins: a diversity of intracellular lipid droplet proteins. Lipids Health Dis. 2017;16(1):83. https://doi.org/10.1186/s12944-017-0473-y
2. Conte M, Franceschi C, Sandri M, Salvioli S. Perilipin 2 and Age-Related Metabolic Diseases: A New Perspective. Trends Endocrinol Metab. 2016;27(12):893-903. https://doi.org/10.1016/j.tem.2016.09.001
3. Sztalryd C, Brasaemle DL. The perilipin family of lipid droplet proteins: Gatekeepers of intracellular lipolysis. Biochim Biophys Acta Mol Cell Biol Lipids. 2017;1862(10 Pt B):1221-1232. https://doi.org/10.1016/j.bbalip.2017.07.009
4. Uniprot (Q99541)
5. Chong BM, Reigan P, Mayle-Combs KD, Orlicky DJ, McManaman JL. Determinants of adipophilin function in milk lipid formation and secretion. Trends Endocrinol Metab. 2011;22(6):211-217. https://10.1016/j.tem.2011.04.003
Alternate Names
ADFP, Adipophilin, Perilipin2, PLIN2
Gene Symbol
PLIN2
UniProt
Additional Perilipin-2 Products
Product Documents for Perilipin-2/ADFP Antibody - BSA Free
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Product Specific Notices for Perilipin-2/ADFP 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.
Related Research Areas
Citations for Perilipin-2/ADFP Antibody - BSA Free
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Application: Western BlotSample Tested: Liver whole cell extract and Liver whole tissue homogenate, Sample Amount: 50ugSpecies: RatVerified Customer | Posted 06/16/2020Perilipin2Concentration 1:1000, over night Secondary antibody: 1: 20000
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Protocols
View specific protocols for Perilipin-2/ADFP Antibody - BSA Free (NB110-40877):
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 4 degrees C.
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 25 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.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- ISH-IHC Protocol for Chromogenic Detection on Formalin Fixed Paraffin Embedded (FFPE) Tissue
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for Perilipin-2/ADFP Antibody - BSA Free
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Q: I went trought your product datasheet, but I didn't find any information about the dilution I have to use for the western blot and if it's in 5% milk-TBST or other blocking buffer. Could you please provide me those information?
A: For our product NB110-40877, we recommend a starting dilution of 2 ug/ml for western blot. Our recommended diluent buffer is 5% nonfat milk in TBST.
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Q: I would appreciate if you are able to provide the exact amino acid sequence of the immunogens. The customer would like to check the percentage alignment with the chicken protein to determine which of the antiodies woudl be better to try in this species.
A: I am unable to provide you with this information however I can tell you that there is a 71% homology between the chicken protein and human protein. Our lab only recommends use if the homology is 85% or above.
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Q: I went trought your product datasheet, but I didn't find any information about the dilution I have to use for the western blot and if it's in 5% milk-TBST or other blocking buffer. Could you please provide me those information?
A: For our product NB110-40877, we recommend a starting dilution of 2 ug/ml for western blot. Our recommended diluent buffer is 5% nonfat milk in TBST.
-
Q: I would appreciate if you are able to provide the exact amino acid sequence of the immunogens. The customer would like to check the percentage alignment with the chicken protein to determine which of the antiodies woudl be better to try in this species.
A: I am unable to provide you with this information however I can tell you that there is a 71% homology between the chicken protein and human protein. Our lab only recommends use if the homology is 85% or above.
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