Invitrogen

IL-6 Recombinant Rabbit Monoclonal Antibody (4H16L21)

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IL-6 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of IL-6 was performed using 70% confluent log phase HUVEC cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with IL-6 Recombinant Rabbit Monoclonal Antibody (4H16L21) (Product # 701028) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly Golgi Complex and Endoplasmic Reticulum localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

IL-6 Antibody in Western Blot (WB) — Western blot was performed using Anti-IL-6 Recombinant Rabbit Monoclonal Antibody (4H16L21) (Product # 701028) and a 24 kDa band corresponding to IL-6 was observed across upon treatment of HUVEC cells with LPS and Brefeldin A. Whole cell extracts (30 µg lysate) of HUVEC (Lane 1) and HUVEC treated with LPS and Brefeldin A (0.5 µg/mL LPS for 24 h and 300 ng/mL Brefeldin A for last 20 h of LPS stimulation) (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).Expression of IL-6 was observed to be to be upregulated upon treatment of HUVEC cells with LPS and Brefeldin A. An uncharacterized band (*) was observed at ~55 kDa.

IL-6 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of IL-6 on PHA-M stimulated THP-1 cells with or without LPS treatment. Cells were fixed with 4% formaldehyde for 30 min on ince and permeabilized with IC permeabilization buffer (Product # PB001). After incubation with blocking buffer (Product # 37525) for 30 min on ice, cells were then stained with anti-IL-6 rabbit monoclonal antibody (Product # 701028) at indicated dilution with IC permeabilization buffer for 30 min on ice. After washing with ice-cold IC permeabilization buffer for 3 times, the cells were stained with DyLight 488 goat anti-rabbit secondary antibody (Product # 35552) for 30 min on ice. A representative 10,000 cells were acquired for each sample.

IL-6 Antibody in ELISA (ELISA) — Indirect ELISA was performed using various dilutions of Anti-IL-6 Rabbit Monoclonal Antibody (Product # 701028) to detect IL-6 recombinant protein coated onto the plate. A non-linear regression analysis was performed (4 PL) and LOD and LOQ for the antibody was determined.

Product Details

701028

Applications	Tested Dilution	Publications
Western Blot (WB)	0.1-1 µg/mL	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	1:200	-
Flow Cytometry (Flow)	1:100	-
ELISA (ELISA)	Assay-dependent	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human
Host/Isotype	Rabbit / IgG
Class	Recombinant Monoclonal
Type	Antibody
Clone	4H16L21
Immunogen	proprietary
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2532352

Product Specific Information

701028 was successfully to detect IL-6 in Flow application.

Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Target Information

Interleukin 6 (IL-6) is a multifunctional 26 kD protein originally discovered in the medium of RNA-stimulated fibroblastoid cells. IL-6 appears to be directly involved in the responses that occur after infection and cellular injury, and it may prove to be important as IL-1 and TNF-a in regulating the acute phase response. IL-6 is reported to be produced by fibroblasts, activated T cells, activated monocytes or macrophages and endothelial cells. IL-6 acts upon a variety of cells including myeloid progenitor cells, T cells, B cells, and hepatocytes. In addition, IL-6 appears to interact with IL-2 in the proliferation of T lymphocytes. IL-6 potentiates the proliferative effect of IL-3 on multipotential hematopoietic progenitors. IL-6 plays a critical role in B-cell differentiation to plasma cells and is a potent growth factor for plasmacytoma and myeloma. IL-6 is a very useful culture supplement for the generation of a high number of antibody-producing hybridomas. Primarily produced at sites of acute and chronic inflammation, IL-6 is secreted into the serum and induces a transcriptional inflammatory response through interleukin 6 receptor, alpha. The functioning of IL-6 is implicated in a wide variety of inflammation-associated disease states including diabetes mellitus and systemic juvenile rheumatoid arthritis.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Effect of PM2.5 on invasion and proliferation of HeLa cells and the expression of inflammatory cytokines IL-1 and IL-6.

Oncology letters

Huang K,Li W,Chen Y,Zhu J

Published figure using IL-6 recombinant monoclonal antibody (Product # 701028) in ELISA

Sat Dec 01 00:00:00 EST 2018

Four assay designs and on-chip calibration: gadgets for a sepsis protein array.

Analytical chemistry

Buchegger P,Preininger C

701028 was used in ELISA to develop four protein microarray assays for the early diagnosis of sepsis based on the measurement of CRP, PCT and IL-6

Tue Mar 18 00:00:00 EDT 2014

Bioinformatics

Protein Aliases: B-cell differentiation factor; B-cell hybridoma growth factor; B-cell stimulatory factor 2; BSF-2; CDF; CTL differentiation factor; H-IL-6; Hybridoma growth factor; IFN-beta-2; IL-6; ILN; Interferon beta-2; Interleukin; interleukin BSF-2; Interleukin HP-1; Interleukin-6; Interleukin6; M-IL-6; R-IL-6

Gene Aliases: BSF-2; BSF2; CDF; HGF; HSF; IFN-beta-2; IFNB2; IL-6; IL6

UniProt ID: (Human) P05231, (Mouse) P08505

Entrez Gene ID: (Human) 3569, (Mouse) 16193

Function(s)

Process(es)

cytokine activity interleukin-6 receptor binding protein binding growth factor activity receptor binding

IL-6 Recombinant Rabbit Monoclonal Antibody (4H16L21)

Product Details

701028

Western Blot (WB)

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

ELISA (ELISA)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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