Recombinant Anti-IL-6 antibody [EPR20653] - BSA and Azide free (ab245770)
![Western blot - Anti-IL-6 antibody [EPR20653] - BSA and Azide free (ab245770)](https://www.abcam.com/ps/products/245/ab245770/Images/ab245770-446575-anti-il-6-antibody-epr21711-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] - BSA and Azide free (ab245770)")
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 - BSA and Azide free
- Suitable for: ICC/IF, IP, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IL-6 antibody [EPR20653] - BSA and Azide free
See all IL-6 primary antibodies -
Description
Rabbit monoclonal [EPR20653] to IL-6 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, WBmore 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
- ICC/IF: HUVEC treated with Lipopolysaccharides (LPS) (0.5 µg/ml 24h) and Brefeldin A (BFA) (300 ng/ml 20h) cells; Wild-type A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 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
ab245770 is the carrier-free version of ab214429.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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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Conjugation kits
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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 ab245770 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 |
|---|---|---|
| ICC/IF |
Use at an assay dependent concentration.
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| IP |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Detects a band of approximately 21-28 kDa (predicted molecular weight: 23 kDa).
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| Notes |
|---|
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ICC/IF
Use at an assay dependent concentration. |
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IP
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Detects a band of approximately 21-28 kDa (predicted molecular weight: 23 kDa). |
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] - BSA and Azide free (ab245770)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)This data was developed using the same antibody clone in a different buffer formulation (ab214429).
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] - BSA and Azide free (ab245770)](https://www.abcam.com/ps/products/245/ab245770/Images/ab245770-337927-anti-il-6-antibody-epr20653-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR20653] - BSA and Azide free (ab245770)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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab214429).
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![Immunoprecipitation - Anti-IL-6 antibody [EPR20653] - BSA and Azide free (ab245770)](https://www.abcam.com/ps/products/245/ab245770/Images/ab245770-337928-anti-il-6-antibody-epr20653-immunoprecipitation.jpg)
Immunoprecipitation - Anti-IL-6 antibody [EPR20653] - BSA and Azide free (ab245770)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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab214429).
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![Anti-IL-6 antibody [EPR20653] - BSA and Azide free (ab245770)](https://www.abcam.com/ps/products/245/ab245770/Images/ab245770-5-benefits-of-recombinant-antibodies.png)
Anti-IL-6 antibody [EPR20653] - BSA and Azide free (ab245770)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab245770 has been referenced in 1 publication.
- Wang GC et al. Inflammation induced by lipopolysaccharide advanced androgen receptor expression and epithelial-mesenchymal transition progress in prostatitis and prostate cancer. Transl Androl Urol 10:4275-4287 (2021). PubMed: 34984192
