IL-6 Antibody - Azide and BSA Free

Novus Biologicals | Catalog # NB600-1131

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Monkey

Cited:

Human, Mouse, Rat, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Knockdown Validated

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, IF/IHC, Knockdown Validated

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

Azide and BSA Free
View all IL-6 Primary Antibodies »

Product Specifications

Immunogen

This whole rabbit serum was prepared by repeated immunizations with recombinant human IL-6 produced in E.coli. (Uniprot: P05231)

Reactivity Notes

Use in Rat reported in scientific literature (PMID:33651836).

Localization

Secreted

Specificity

Anti-IL-6 antiserum detects recombinant and native IL-6 present in body fluids and cell supernatants in various assays (ie. IL-1 stimulated IL-6 production from fibroblasts).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for IL-6 Antibody - Azide and BSA Free

Western Blot: IL-6 Antibody [NB600-1131]

Western Blot: IL-6 Antibody [NB600-1131]

Western Blot: IL-6 Antibody [NB600-1131] - Western Blot: IL-6 Antibody [HRP] [NBP1-42762] - Lane 1: Human IL-6. Lane 2: none. Load: 50 ng per lane. Primary antibody: none. Secondary antibody: Peroxidase Human IL-6 secondary antibody at 1:1,000 for 60 min at RT. Block: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 20 kDa for Human IL-6. Other band(s): none. Image using the HRP form of this antibody.
Immunohistochemistry: IL-6 Antibody [NB600-1131]

Immunohistochemistry: IL-6 Antibody [NB600-1131]

IL-6-Antibody-Immunocytochemistry-Immunofluorescence-NB600-1131-img0015.jpg
Western Blot: IL-6 Antibody [NB600-1131]

Western Blot: IL-6 Antibody [NB600-1131]

Western Blot: IL-6 Antibody [NB600-1131] - Protein was resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose. The blot shows detection of a band 21 kDa in size corresponding to anti-IL6 antibody. Molecular weight markers are also shown (MW). After transfer, the membrane was blocked for 30 minutes with 1% BSA-TBST. Detection occurred using peroxidase conjugated anti-Rabbit IgG secondary antibody diluted 1:40,000 in blocking buffer for 30 min at RT followed by reaction with FemtoMax chemiluminescent substrate.
IL-6 Antibody

IL-6 Antibody

Western Blot of IL-6 Antibody. Lane 1: Opal Prestained Molecular Weight Marker
IL-6 Antibody

Immunocytochemistry/ Immunofluorescence: IL-6 Antibody [NB600-1131] -

Immunocytochemistry/ Immunofluorescence: IL-6 Antibody [NB600-1131] - GBM ECs express IL-6. a Human brain ECs were treated with glioma-CM for 24 h, & cell lysates were subjected to multiplex cytokine array analysis. Left, a representative blot. Right, quantified dot intensity of most significantly changed cytokines. b Human microvascular brain ECs were treated with glioma-CM that were harvested from different human glioma cells. Cell lysates were immunoblotted. c Human microvascular brain ECs & tumor-associated ECs isolated from different GBM patients were subjected to immunoblot analysis. d Mouse GBM was induced by orthotopic injection of GL26 glioma cells into wild-type mouse. The brain sections that include normal brains & tumors were stained with anti-CD31, anti-IL-6, & anti-CSF-1 antibodies. Representative immunofluorescence images are shown. Right, enlarged area in normal & tumor tissues. Bar represents 50 μm. Zoom-in factor: 4 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29422647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for IL-6 Antibody - Azide and BSA Free

Application
Recommended Usage

ELISA

1:1000-1:5000

Immunocytochemistry/ Immunofluorescence

1:10 - 1:500

Immunohistochemistry

1:400-1:800

Immunohistochemistry-Paraffin

1:400 - 1:800

Immunoprecipitation

1:400-1:800

Western Blot

1:500-1:2000
Application Notes
This product has been tested in western blot. This antibody is suitable for use in neutralizations, ELISA, radioimmunoassay, and immunoprecipitation. Reactivity in other immunoassays is unknown. Expect ~23.7kDa. In Western blot analysis of natural cell products or human body fluids, multiple bands of IL-6 will appear due to the variable amount of glycosylation on the molecule. For immunoblot use the supernatants or lysates of 2 x 106 endotoxin-stimulated human peripheral blood mononuclear cells (PBMC). PBMC are stimulated for 24 hours with 1% (v/v) human serum plus 10ng/mL E. coli LPS. For immunoprecipitation, pre-clearing with a non-specific rabbit IgG is helpful to reduce background. The antiserum is useful for neutralization of human of IL-6 activity in bioassays. For neutralization, incubate the sample with a 1:400 dilution of the antiserum for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG (heat inactivated) is recommended. In neutralization experiments in vitro, this antibody does not result in enhanced activity of IL-6. However, because antibodies to IL-6 may act as a soluble receptor in vivo, some antibodies to IL-6 act as carriers and enhance IL-6 activity.

Reviewed Applications

Read 1 review rated 1 using NB600-1131 in the following applications:

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

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

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: IL-6

The antiserum detects recombinant and native IL-6 present in body fluids and cell supernatants in various assays (ie. IL-1 stimulated IL-6 production from fibroblasts). In Western blot analysis of natural cell products or human body fluids, multiple bands of IL-6 will appear due to the variable amount of glycosylation on the molecule (21-28 kDa). The antiserum is also useful for neutralization of human IL-6 activity in bioassays. For neutralization, incubate the sample with a 1:400 dilution of the antiserum for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG (heat inactivated) is recommended. In neutralization experiments in vitro, this antibody does not result in enhanced activity of IL-6. However, because antibodies to IL-6 may act as a soluble receptor in vivo, some antibodies to IL-6 act as carriers and enhance IL-6 activity.

Long Name

Interleukin 6

Alternate Names

BSF-2, BSF2, IFNB2, IL6, MGI-2A

Entrez Gene IDs

3569 (Human); 3569 (Human)

Gene Symbol

IL6

UniProt

P05231

Additional IL-6 Products

Product Documents for IL-6 Antibody - Azide and BSA Free

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 IL-6 Antibody - Azide and 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 IL-6 Antibody - Azide and BSA Free

Customer Reviews for IL-6 Antibody - Azide and BSA Free (1)

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  • IL-6 Antibody
    Name: Lenka Uherková
    Application: Simple Western
    Sample Tested: skin wound - healthy
    Species: Rat
    Verified Customer | Posted 02/12/2020
    IL-6 Antibody - Azide and BSA Free NB600-1131
    Bio-Techne Response
    This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.

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Protocols

View specific protocols for IL-6 Antibody - Azide and BSA Free (NB600-1131):


Materials

1) 1 Phosphate buffered saline (pH 7.6): NaCl 137mmol/L, KCl 2.7mmol/L, Na2HPO4 4.3mmol/L, KH2PO4 1.4 mmol/L
2) Citrate buffer, 0.01 M, pH6.0, Sodium Citrate 3g, Citric acid 0.4g
3) 3% Hydrogen peroxide
4) Primary antibody
5) Blocking serum (normal serum)
6) Biotinylated secondary antibody
7) DAB staining kit

Methods

1. Dewax and hydration of slides using xylene and EtOH:
Dry slides for 20 min in a 60 C oven
Add Xylene, 2 x 10 min
100%, 95%, 80%, and 70% EtOH, 5 min each EtOH concentration
Rinse in PBS, 5'

2 Antigen retrieval method (only for paraffin slides)
1a. High-pressure antigen retrieval procedure (recommended method)
Place slides in a glass slide holder (ensure that the slide holder is completely filled with slides, slides without sections if necessary, to ensure even heating. The entire slide holder is immersed in 1000 ml of Citrate buffer (0.01M, pH6.0) within a pressure cooker
Once steam is produced, and ONLY when steam is visible, from the pressure cooker (usually 15-20 min), the required high-pressure will have been reached, and slides will be incubated for 2 min.
Turn off heat, and allow buffer and slides to cool to room temperature
Slides are then rinsed in PBS for 5 minutes
2. Add 3% hydrogen peroxide solution, 10'at RT, then PBS, 3X5'
3. Normal blocking serum, 20'at RT
4. Incubate with Primary Ab, 4C overnight or 1.5 hours at 37C
5. Rinse with PBS, 3 X 5' each rinse
6. Add Biotin-conjugated second antibody, 10'at RT
7. Rinse with PBS, 3 X 5' each rinse
8. Add Streptavidin-Peroxidase, 10'at RT
9. Rinse with PBS, 3 X 5' each rinse
10. Staining with DAB solution, 2-5'under microscope
11. Stop the reaction by washing in tap water
12. Counterstain in Haematoxylin for 3-5 minutes
13. 75%, 80%, 95% and 100% ethanol, 5x2', xylene 2 x 10'

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FAQs for IL-6 Antibody - Azide and BSA Free

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    • Q: I had a quick question regarding NB600-1131SS IL-6 antibody. Not sure if you guys would be able to answer but how low do you think I could go on protein amount loaded to pick up a band? I see on the data sheet you guys have loaded 50ug and I cant load that much based on the nature of my samples.
      A: We usually recommend anywhere from 10-50 ug/lane of protein. I wouldn't recommend using much lower than 10 ug of sample.

    • Q: I was wondering if you know of any blocking antibodies against IL-6 or VEGF-C that can be used in cell culture to quench these cytokines?
      A: We have no antibodies that are currently known to show blocking antibodies to VEGF-C. Please see this link to our IL-6 antibodies with blocking/neutralizing activity can be found at this link.
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