Ultra-LEAF™ Purified anti-human CD274 (B7-H1, PD-L1) Antibody

Product Details

Verified Reactivity

Human

Reported Reactivity

African Green, Baboon, Cynomolgus, Rhesus

Antibody Type

Monoclonal

Host Species

Mouse

Immunogen

Full length human PD-L1

Formulation

0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.

Endotoxin Level

Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.

Preparation

The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.

Concentration

The antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. Older lots may have also been bottled at 1 mg/mL. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.

Application

FC - Quality tested
IHC, Block - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤2.0 µg per million cells in 100 µl volume or 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clone 29E.2A3 is reported to recognize an epitope on PD-L1 within the PD-L1-CD80 binding region⁵. Additional reported applications (for the relevant formats) include: blocking^1-3 and immunohistochemical staining of acetone-fixed frozen sections¹. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 329715, 329716, 329745 - 329748). 

It has been observed that clone 29E.2A3 is able to bind to Alexa Fluor® 700 antibody conjugates during multi-color immunofluorescent staining. This interaction can be resolved by sequentially staining with the 29E.2A3 antibody first and then followed by the Alexa Fluor® 700 conjugate of interest.

Clone 29E.2A3 does not work in Western blot applications⁷.

Application References

1. Brown J, et al. 2003. J. Immunol. 170:1257. (FC, IHC, Block)
2. Radziewicz H, et al. 2007. J. Virol. 81:2545. (Block)
3. Nakamoto N, et al. 2009. PLoS Pathog. 5:e1000313. (Block)
4. Barsoum IB, et al. 2014. Cancer Res. 74:665. PubMed
5. Haile, S et al. 2013. J. Immunol. 191:2829.
6. RL M, et al. 2015. PNAS. 112:6506-6514. PubMed
7. Mahoney KM, et al. 2015. Cancer Immunol. Res. 3:1308.

Product Citations

1. Zhang Z, et al. 2022. Cancer Gene Ther. 29:722. PubMed
2. Yang Z, et al. 2022. Cancer Discov. 12:1942. PubMed
3. Nishi W, et al. 2023. Nat Commun. 14:3157. PubMed
4. Wang H, et al. 2020. J Exp Clin Cancer Res. 39:29. PubMed
5. Rieder SA, et al. 2020. Cell Mol Immunol. . PubMed
6. Baleeiro RB, et al. 2022. Oncoimmunology. 11:2080329. PubMed
7. Curnock AP, et al. 2021. JCI Insight. 6:. PubMed
8. Meng Q, et al. 2021. J Immunother Cancer. 9:. PubMed

RRID

AB_11149486 (BioLegend Cat. No. 329715)
AB_11149486 (BioLegend Cat. No. 329716)
AB_11149486 (BioLegend Cat. No. 329745)
AB_11149486 (BioLegend Cat. No. 329746)
AB_11149486 (BioLegend Cat. No. 329747)
AB_11149486 (BioLegend Cat. No. 329748)

Antigen Details

Distribution

T cells, B cells, NK cells, monocytes/macrophages, granulocytes and dendritic cells

Function

CD274 is involved in the costimulatory signal, essential for T lymphocyte proliferation and production of IL-10 and IFN-γ, in an IL-2-dependent and a PD-1-CD1-independent manner. Its interaction with PD-1-CD1 inhibits T-cell proliferation and cytokine production.

Ligand/Receptor

PD-1 (PDCD1)

Cell Type

B cells, Dendritic cells, Fibroblasts, Granulocytes, Macrophages, Monocytes, NK cells, T cells

Biology Area

Cancer Biomarkers, Costimulatory Molecules, Immunology

Molecular Family

Adhesion Molecules, CD Molecules, Immune Checkpoint Receptors

Antigen References

1. Sharpe A, et al. 2007. Nat. Immunol. 8:239.

Gene ID

29126 View all products for this Gene ID

UniProt

View information about CD274 on UniProt.org

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Material Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

Related FAQs

Do you guarantee that your antibodies are totally pathogen free?

BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.

Does BioLegend test each Ultra-LEAF™ antibody by functional assay?

No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.

Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?

No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.

Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?

We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.

Other Formats

Certificate of Analysis