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Invitrogen™ IL-12 p35 Monoclonal Antibody (4D10p35), eFluor™ 660, eBioscience™, Invitrogen™

Catalog No. 501124452
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Catalog No. 50-112-4452 Supplier Invitrogen™ Supplier No. 50735280
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Rat Monoclonal Antibody

Description: This 4D10p35 monoclonal antibody reacts with mouse p35, a component of two of the four members of the interleukin (IL)-12 family of cytokines. The IL-12 family members are heterodimeric cytokines composed of an alpha-chain, such as p35, and a beta-chain, such as p40. The alpha-chain proteins are thought to be the limiting factors for secretion of IL-12 family member cytokines, as expression of the alpha-chain is more restricted than that of the beta-chain proteins, and the two subunits may need to be co-expressed in order for cytokine secretion to occur. IL-12 family members are secreted primarily by antigen-presenting cells in response to various TLR agonists and other inflammatory stimuli. The exact types of stimuli present and kinetics of stimulation determine the balance of IL-12 family member production. The p35 protein was first identified as part of the IL-12 cytokine, which is also known as IL-12p70. Secretion is induced by IFN gamma and agonists of TLR3, 4, or 7 and is inhibited by Th2 cytokines. IL-12 is essential for the differentiation and maintece of Th1 cells. The other subunit, p40, is expressed in much higher quantities than p35, and may be secreted independently as a monomer or homodimer. Free p40 may act as an antagonist of IL-12 function. More recently, p35 was also found to dimerize with EBI3 (Epstein-Barr Virus-Induced Gene 3) to form IL-35.

Interleukin 12A is a novel signaling cytokine belonging to the Interleukin-6 protein superfamily. Interleukin-12 alpha chain precursor (IL-12A) is a 35-kD subunit of disulfide-linked heterodimer of cytokine IL-12 and has a broad array of biological activities. Reports suggest that IL-12A can act as a growth factor for activated T and NK cells and enhance the lytic activity of NK/lymphokine-activated Killer cells. IL-12A stimulates the production of IFN-gamma by resting PBMC as well as T-cell-independent induction of interferon (IFN)-gamma, and helps in the differentiation of both Th1 and Th2 cells. It is also involved in positive regulation of tyrosine phosphorylation of Stat4 protein. The signaling process of IL-12A in innate immunity requires nitric oxide synthase 2A. Along with EBI3, IL-12A forms a novel heterodimeric cytokine, designated as interleukin-35 (IL-35), that may be specifically produced by T reg cells and is required for maximal suppressive activity.
TRUSTED_SUSTAINABILITY

Specifications

Antigen IL-12 p35
Applications Flow Cytometry
Classification Monoclonal
Clone 4D10p35
Concentration 0.2 mg/mL
Conjugate eFluor 660
Formulation PBS with 0.09% sodium azide; pH 7.2
Gene IL12A
Gene Accession No. P43431
Gene Alias CLMF; CLMF p35; cytotoxic lymphocyte maturation factor 1, p35; cytotoxic lymphocyte maturation factor 35 kDa subunit; il 12; IL-12 subunit p35; IL-12, subunit p35; Il12a; IL-12A; IL12p35; IL-12p35; IL35 subunit; ILN; Interleukin; Interleukin 12; interleukin 12 p35 subunit; interleukin 12, p35; interleukin 12a; interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocyte maturation factor 1, p35); Interleukin12; interleukin-12 alpha chain; interleukin-12 beta p35 subunit; interleukin-12 p35 subunit; interleukin-12 subunit alpha; Ll12a; NF cell stimulatory factor chain 1; NFSK; NK cell stimulatory factor chain 1; NKSF1; P35
Gene Symbols IL12A
Host Species Rat
Immunogen Mouse IL-35-Fc recombinant protein.
Purification Method Affinity chromatography
Quantity 25 μg
Regulatory Status RUO
Primary or Secondary Primary
Gene ID (Entrez) 16159
Target Species Mouse
Content And Storage 4°C, store in dark, DO NOT FREEZE!
Product Type Antibody
Form Liquid
Isotype IgG2a κ
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I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

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