Recombinant Anti-Pellino 1 antibody [EPR19302] (ab199336)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19302] to Pellino 1
- Suitable for: WB, IP, ICC/IF
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Pellino 1 antibody [EPR19302]
See all Pellino 1 primary antibodies -
Description
Rabbit monoclonal [EPR19302] to Pellino 1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human PELI1 full length recombinant protein; Human fetal liver, fetal heart and fetal kidney lysates; PC-12, NIH/3T3, Ramos, HeLa, RAW 264.7 (treated with 1 µg/ml LPS for 6 hours) and PC-12 (treated with 1 µg/ml LPS for 6 hours) whole cell lysates. ICC/IF: HeLa and Ramos cells. IP: Ramos whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19302 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab199336 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 |
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WB |
1/1000. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa).
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IP |
1/30.
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ICC/IF |
1/100.
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Notes |
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WB
1/1000. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa). |
IP
1/30. |
ICC/IF
1/100. |
Target
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Function
Scaffold protein involved in the IL-1 signaling pathway via its interaction with the complex containing IRAK kinases and TRAF6. Required for NF-kappa-B activation and IL-8 gene expression in response to IL-1. -
Sequence similarities
Belongs to the pellino family. - Information by UniProt
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Database links
- Entrez Gene: 57162 Human
- Entrez Gene: 67245 Mouse
- Entrez Gene: 305549 Rat
- Omim: 614797 Human
- SwissProt: Q96FA3 Human
- SwissProt: Q8C669 Mouse
- Unigene: 727530 Human
- Unigene: 28957 Mouse
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Alternative names
- E3 ubiquitin-protein ligase pellino homolog 1 antibody
- PELI1 antibody
- PELI1_HUMAN antibody
see all
Images
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All lanes : Anti-Pellino 1 antibody [EPR19302] (ab199336) at 1/5000 dilution
Lane 1 : Human PELI1 full length recombinant protein
Lane 2 : Human PELI2 full length recombinant protein
Lane 3 : Human PELI3 full length recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Human PELI 1 full length recombinant protein contain aa1-418 with a His-Tag®. Human PELI 2 full length recombinant protein contain aa1-420 with a His-Tag®. Human PELI3 full length recombinant protein contain aa1-469 with a His-Tag®. All three recombinant human fragment proteins were made in-house.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Pellino 1 with ab199336 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [EPR19302] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab199336 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution. -
Lane 1 (input): Mouse brain lysate, 10μg
Lane 2 (+): Mouse brain lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab199336 in Mouse brain lysateAb199336 immunoprecipitating Pellino in Mouse brain lysate. For western blotting, primary antibody used was ab199336 at 1:500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST
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All lanes : Anti-Pellino 1 antibody [EPR19302] (ab199336) at 1/1000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Pellino 1 antibody [EPR19302] (ab199336) at 1/1000 dilution
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Pellino 1 antibody [EPR19302] (ab199336) at 1/1000 dilution
Lane 1 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : Untreated RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 1 µg/ml LPS for 6 hours whole cell lysate
Lane 5 : Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 1 µg/ml LPS for 6 hours whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The expression of Pellino 1 can be induced by LPS. (PMID: 21204785).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (Human Burkitt's lymphoma cell line) cells labeling Pellino 1 with ab199336 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining on Ramos cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [EPR19302] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab199336 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution. -
Pellino 1 was immunoprecipitated from 1mg of Ramos (Human Burkitt's lymphoma cell line) whole cell lysate with ab199336 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199336 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Ramos whole cell lysate 10µg (Input).
Lane 2: ab199336 IP in Ramos whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR19302] - Isotype Control (ab172730) instead of ab199336 in Ramos whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: (A): 30 seconds; (B): 3 minutes.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (7)
ab199336 has been referenced in 7 publications.
- Liu Y et al. Insulin Resistance and Pellino-1 Mediated Decrease in the Activities of Vasodilator Signaling Contributes to Sunitinib-Induced Hypertension. Front Pharmacol 12:617165 (2021). PubMed: 33841146
- Wei W et al. IL-1 signaling pathway molecules as key markers in childhood asthma. Pediatr Allergy Immunol 32:305-313 (2021). PubMed: 33025692
- Luo S et al. MiR-301a-3p Advances IRAK1-Mediated Differentiation of Th17 Cells to Promote the Progression of Systemic Lupus Erythematosus via Targeting PELI1. J Healthc Eng 2021:2982924 (2021). PubMed: 34931135
- Kinsella S et al. Increased A20-E3 ubiquitin ligase interactions in bid-deficient glia attenuate TLR3- and TLR4-induced inflammation. J Neuroinflammation 15:130 (2018). PubMed: 29720226
- Yuan Y et al. MicroRNA High Throughput Loss-of-Function Screening Reveals an Oncogenic Role for miR-21-5p in Hodgkin Lymphoma. Cell Physiol Biochem 49:144-159 (2018). PubMed: 30184526
- Choi SW et al. PELI1 Selectively Targets Kinase-Active RIP3 for Ubiquitylation-Dependent Proteasomal Degradation. Mol Cell 70:920-935.e7 (2018). PubMed: 29883609
- Wang H et al. PELI1 functions as a dual modulator of necroptosis and apoptosis by regulating ubiquitination of RIPK1 and mRNA levels of c-FLIP. Proc Natl Acad Sci U S A 114:11944-11949 (2017). PubMed: 29078411