Recombinant Anti-IL-6 antibody [EPR20653] (ab214429)
![Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)](https://www.abcam.com/ps/products/214/ab214429/Images/ab214429-446564-anti-il-6-antibody-epr20653-western-blot-mps-a549-wt-brefeldin-a-5ugml-during-the-last-4-h-mps-a549-wt-il1b.jpg "Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20653] to IL-6
- Suitable for: WB, ICC/IF, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IL-6 antibody [EPR20653]
See all IL-6 primary antibodies -
Description
Rabbit monoclonal [EPR20653] to IL-6 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HUVEC treated with 0.5 µg/ml Lipopolysaccharides (LPS) for 24 hours and 300 ng/ml Brefeldin A (BFA) for 20 hours whole cell lysate; Wild-type A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate ICC/IF: HUVEC treated with Lipopolysaccharides (LPS) (0.5 µg/ml 24h) and Brefeldin A (BFA) (300 ng/ml 20h) cells. IP: HUVEC treated with 0.5 µg/ml Lipopolysaccharides (LPS) for 24 hours and 300 ng/ml Brefeldin A (BFA) for 20 hours 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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Form
Liquid -
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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20653 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab214429 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 |
1/1000. Detects a band of approximately 21-28 kDa (predicted molecular weight: 23 kDa).
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| ICC/IF |
1/500.
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| IP |
1/30.
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 21-28 kDa (predicted molecular weight: 23 kDa). |
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ICC/IF
1/500. |
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IP
1/30. |
Target
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Function
Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig-secreting cells Involved in lymphocyte and monocyte differentiation. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation Acts on B-cells, T-cells, hepatocytes, hematopoeitic progenitor cells and cells of the CNS. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance. -
Involvement in disease
Genetic variations in IL6 are associated with susceptibility to rheumatoid arthritis systemic juvenile (RASJ) [MIM:604302]. An inflammatory articular disorder with systemic-onset beginning before the age of 16. It represents a subgroup of juvenile arthritis associated with severe extraarticular features and occasionally fatal complications. During active phases of the disorder, patients display a typical daily spiking fever, an evanescent macular rash, lymphadenopathy, hepatosplenomegaly, serositis, myalgia and arthritis.
Note=A IL6 promoter polymorphism is associated with a lifetime risk of development of Kaposi sarcoma in HIV-infected men. -
Sequence similarities
Belongs to the IL-6 superfamily. -
Post-translational
modificationsN- and O-glycosylated. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 3569 Human
- Omim: 147620 Human
- SwissProt: P05231 Human
- Unigene: 654458 Human
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Alternative names
- Interleukin BSF 2 antibody
- B cell differentiation factor antibody
- B cell stimulatory factor 2 antibody
see all
Images
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Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)All lanes : Anti-IL-6 antibody [EPR20653] (ab214429) at 1/1000 dilution
Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate
Lane 2 : Wild-type A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate
Lane 3 : IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate
Lane 4 : IL-6 knockout A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?
Additional bands at: 35 kDa (possible non-specific binding)Lanes 1 - 4: Merged signal (red and green). Green - ab214429 observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab214429 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type and IL-6 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab214429 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR20653] (ab214429)](https://www.abcam.com/ps/products/214/ab214429/Images/ab214429-281320-anti-il-6-antibody-epr20653-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR20653] (ab214429)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC (human umbilical vein endothelial cell line) cells, untreated or treated with Lipopolysaccharides (LPS) (0.5 µg/ml 24 hours) and Brefeldin A (BFA) (300 ng/ml 20 hours), labeling IL6 with ab214429 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing the cytoplasmic expression increased on HUVEC cells treated with Lipopolysaccharides (LPS) (0.5 µg/ml, 24 hours) and Brefeldin A (BFA) (300 ng/ml, 20 hours).
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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![Immunoprecipitation - Anti-IL-6 antibody [EPR20653] (ab214429)](https://www.abcam.com/ps/products/214/ab214429/Images/ab214429-281318-anti-il-6-antibody-epr20653-immunoprecipitation.jpg)
Immunoprecipitation - Anti-IL-6 antibody [EPR20653] (ab214429)IL6 was immunoprecipitated from 0.35 mg of HUVEC (human umbilical vein endothelial cell line) treated with Lipopolysaccharides (LPS) and Brefeldin A (BFA) whole cell lysate with ab214429 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214429 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1: HUVEC treated with 0.5 μg/ml LPS for 24 hours and 300 ng/ml BFA for 20 hours whole cell lysate 10 μg (Input).
Lane 2: ab214429 IP in HUVEC treated with 0.5 μg/ml LPS for 24 hours and 300 ng/ml BFA for 20 hours whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214429 in HUVEC treated with 0.5 μg/ml LPS for 24 hours and 300 ng/ml BFA for 20 hours whole cell lysate (-).
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)](https://www.abcam.com/ps/products/214/ab214429/Images/ab214429-281319-anti-il-6-antibody-epr20653-western-blot.jpg)
Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)All lanes : Anti-IL-6 antibody [EPR20653] (ab214429) at 1/1000 dilution
Lane 1 : Untreated HUVEC (human umbilical vein endothelial cell line) whole cell lysate
Lane 2 : HUVEC treated with 0.5 µg/ml Lipopolysaccharides (LPS) for 24 hours and 300 ng/ml Brefeldin A (BFA) for 20 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 23 kDa
Observed band size: 21-28 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The MW observed is consistent with the literature (PMID:2523818, PMID: 2783321).
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![Anti-IL-6 antibody [EPR20653] (ab214429)](https://www.abcam.com/ps/products/214/ab214429/Images/ab214429-4-benefits-of-recombinant-antibodies.png)
Anti-IL-6 antibody [EPR20653] (ab214429)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab214429 has been referenced in 2 publications.
- Dou B et al. The roles of metastasis-related proteins in the development of giant cell tumor of bone, osteosarcoma and Ewing's sarcoma. Technol Health Care 29:91-101 (2021). PubMed: 33682749
- Tobío A et al. Oncogenic D816V-KIT signaling in mast cells causes persistent IL-6 production. Haematologica 105:124-135 (2020). PubMed: 30948489