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BioLegend Cell Biology and Functional Proteins
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Featuring

Direct-Blot™ HRP conjugated primary antibodies
BioLegend is proud to introduce our Direct-Blot™ Horse Radish Peroxidase (HRP) directly conjugated primary antibodies. Direct-Blot™ HRP conjugated antibodies are carefully developed and tested, so sensitivity is not compromised as compared to a traditional western blot using a conjugated secondary antibody. For convenience, ease of use, and value, Direct-Blot™ is the most straightforward path to results.

Epitope tag antibodies and affinity gels
BioLegend's epitope tag antibodies are extremely specific, highly characterized and generate clear, quantitative results. The antibodies we supply were developed using rigorous screening techniques resulting in antibodies that work successfully in a variety of immunoassays. To complete this family, we also carry antibodies to other commonly used fusion protein partners and molecular tools, including GFP, GST, Cre-recombinase and GAL4.

Mitospy™ Mitochondrial Probes
MitoSpy™ mitochrondrial localization probes are cell-permeant, fluorogenic chemical reagents used for labeling mitochondria of living cells. They can be used for live-cell imaging applications or they can be fixed to facilitate mounting the specimen with antifade for inverted microscopy or confocal imaging applications. MitoSpy™ Orange CMTMRos can be fixed and permeabilized to allow for co-staining with additional antibodies.

Purified anti-T-bet (clone 4B10)

T-bet, also known as T-box transcription factor T-bet, is considered to be a "master regulator" of Th1 lymphoid development controlling the production of the cytokine IFN-γ. T-bet is widely expressed in hematopoietic cells including stem cells, NK cells, B cells, and T cells. T-bet is critical for the control of microbial pathogens, and knockout animals show multiple physiologic and inflammatory features characteristic of asthma. T-bet expression is optimally observed after IL-12 stimulation and can be suppressed by addition of the Th2 cytokine IL-4 or neutralization of IL-12.

Total cell lysate from PBMC (lane 1, 15 µg) and PBMC treated with 5 µg/mL CD3 and 2 µg/mL CD28 (lane 2, 15 µg) were resolved by electrophoresis (4-12% Bis-Tris), transferred to nitrocellulose, and probed with purified anti-T-bet antibody (clone 4B10). Proteins were visualized using an HRP Goat anti-mouse IgG Antibody and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin antibody (clone 2F1-1) was used as a loading control.

Purified anti-Vimentin (clone O91D3)

Vimentin are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is a widely expressed and highly conserved 54 kD protein that is constitutively expressed in mesenchymal cells, endothelial cells lining blood vessels, renal tubular cells, macrophages, neutrophils, fibroblasts, and leukocytes1,2. Vimentin is used as a marker of mesenchymal cells to distinguish them from epithelial cells3. Increased vimentin expression is frequently used as an EMT marker in cancer4. Autoantibodies to vimentin are commonly found in patients with autoimmune diseases such as Lupus5 and rheumatoid arthritis6, and also found after transplantation7.

HeLa cells were stained with purified anti-Vimentin (clone O91D3) antibody, followed by staining with Alexa Fluor® 594 conjugated goat anti-mouse IgG antibody (red). Actin filaments were labeled with Alexa Fluor® 647 Phalloidin (green). Nuclei were counterstained with DAPI (blue). The image was captured with a 40X objective.

Purified anti-Oct4 (Oct3) (clone 3A2A20)

Oct4 (Octamer binding transcription factor 4) is an important marker of the undifferentiated state and central regulator of pluripotency in ES cells. When embryonic stem cells are triggered to differentiate, Oct4 is downregulated, thus providing a model for the early events linked to somatic differentiation in the developing embryo. Oct4 is a nuclear protein, with a predicted molecular weight of 39 kD.

Western blot analysis of NTERA-2 (lane 1), NF-1 (lane 2) and Hela (lane 3) cells using anti-Oct4 (Oct3) antibody (3A2A20). β-actin (poly6221) was used a loading control.

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