Skip to main content

Key Product Details

Species Reactivity

Human

Applications

ELISA, Flow Cytometry, Neutralization, Surface Plasmon Resonance, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Kappa Clone # 3F12.2D3

Format

Azide and BSA Free

Concentration

1 mg/ml

Product Summary

Immunogen

3F12 was prepared by hyperimmunizing BALB/c mice with purified recombinant human LTalpha expressed in E. coli - the hybridoma clone was selected for its ability to bind LTalpha3 by ELISA. The CDRs of the antibody were grafted to the framework regions of the mice mAb 2G7 to create the chimeric version of the antibody.

Specificity

3F12 binds to human LTalpha3 and LT-alpha-beta (with a KD of ~0.3 nM, determined by BIACORE; ~ 37 pM, determined by ELISA) in both its soluble homotrimeric and membrane heterotrimeric forms. LTalpha belongs to the tumor necrosis factor (TNF) superfamily and is secreted as a homo-trimer (LTalpha3), or is expressed on the cell surface in complex with LTbeta. Lymphotoxin is produced by lymphocytes and in its homotrimeric form binds to TNFRSF1A/TNFR1, TNFRSF1B/TNFBR and TNFRSF14/HVEM. LTalpha3 signaling induces target cells to upregulate many chemokines and cytokines in an NFkB-dependent manner.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Kappa

Scientific Data Images

Western Blot: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) - Chimeric [NBP2-81105] - Western Blot using Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) [NBP2-81105]. Lipopolysaccharide(LPS)-treated human peripheral blood monocyte (PBM) lysate (35ug protein in RIPA buffer) was resolved on a 15% SDS PAGE gel and blots probed with the rabbit IgG-chimeric version of 3F12.2D3 [NBP2-81105] at 2 ug/ml before detection using an anti-rabbit secondary antibody. Protein was detected by chemiluminescence. The expected unmodified running size for Lymphotoxin alpha is 22.3kDa, though the form shown here is likely glycosylated. NBP2-81105 successfully detected Lymphotoxin alpha in LPS-treated human PBM samples.
Flow Cytometry: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) - Chimeric [NBP2-81105] - Flow-cytometry using the Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) [NBP2-81105]. U937 cells were fixed using 2% PFA, permeabilised using 0.5% Triton and stained with unimmunized rabbit IgG antibody (isotype control, black line) or the rabbit IgG-chimeric version of 3F12.2D3 (NBP2-81105, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Applications

Application
Recommended Usage

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Surface Plasmon Resonance

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.

Neutralization

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG2b format, for improved compatibility with existing reagents, assays and techniques. The properties of the chimieric 3F12 is equivalent to that of the original 3F12 mAb. 3F12 is able to bind to Th2 cells and additionally is can functionally block human LTalpha3 versus an isotype control (trastuzumab) - LTalpha3-induced ICAM upregulation is blocked on HUVEC (Human Umbilical Endothelial Cells) (IC50 0.04 nM) and LTalpha3-induced IL-8 is blocked in A549 cells. The antibody has also been used in ELISA studies to detect LT-alpha and to measure binding.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

0.02% Proclin 300

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.

Background: Lymphotoxin-alpha/TNF-beta

TNF-beta (tumor necrosis factor beta, lymphotoxin alpha, TNFSF1, TNF ligand superfamily member 1) a member of the TNF ligand superfamily and a potent lymphoid factor, is mainly produced by activated T and B lymphocytes and leukocytes. TNF-beta is also produced by fibroblasts, astrocytes, myeloma cells, endothelial cells and epithelial cells. TNF-beta mediates inflammatory and immune responses, has cytotoxic activity on tumor cells and other target cells and shares many of the same bioactivities as TNF alpha, including regulation of cell proliferation, differentiation and apoptosis. TNF-beta is not species-specific and human TNF-beta is active on murine cells with a slightly reduced specific activity.

Long Name

Tumor Necrosis Factor beta

Alternate Names

LTA, Lymphotoxin alpha, Lymphotoxinalpha, TNF-beta, tnfb, TNFSF1, TNFSF1B

Gene Symbol

LTA

Product Documents

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices

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.

Loading...