Mouse Anti-CD274 Recombinant Antibody (
9B5F4) (V2LY-0725-LY512)

Name: Mouse Anti-CD274 Recombinant Antibody (9B5F4)
SKU: V2LY-0725-LY512
Rating: 5 (1 reviews)

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Tested Data

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in WB

Western Blot
Positive WB detected in: A549 whole cell lysate, PC-3 whole cell lysate, HepG2 whole cell lysate, MCF-7 whole cell lysate, Hela whole cell lysate
All lanes: PD-L1 antibody at 1:2500
Secondary
Goat polyclonal to Mouse IgG at 1/10000 dilution
Predicted band size: 34, 21 kDa
Observed band size: 55 kDa

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in WB

Western Blot
Positive WB detected in: Hela whole cell lysate
All lanes: PD-L1 antibody at 1:1000, 1:2000, 1:4000
Secondary
Goat polyclonal to Mouse IgG at 1/10000 dilution
Predicted band size: 34, 21 kDa
Observed band size: 55 kDa

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in IHC

IHC image of V2LY-0725-LY512 diluted at 1:100 and staining in paraffin-embedded human tonsil tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in IHC

IHC image of V2LY-0725-LY512 diluted at 1:100 and staining in paraffin-embedded human colon cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in IF

Immunofluorescence staining of 293 cells with V2LY-0725-LY512 at 1:100, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in IF

Immunofluorescence staining of A549 cells with V2LY-0725-LY512 at 1:100, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in IF

Immunofluorescence staining of Hela cells with V2LY-0725-LY512 at 1:100, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in FC

Overlay histogram showing 293 cells stained with V2LY-0725-LY512 (red line) at 1:150. The cells were incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of>10,000 events was performed.

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in FC

Overlay histogram showing A549 cells stained with V2LY-0725-LY512 (red line) at 1:150. The cells were incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of>10,000 events was performed.

Mouse Anti-CD274 Recombinant Antibody (9B5F4) (V2LY-0725-LY512) in FC

Overlay histogram showing Hela cells stained with V2LY-0725-LY512 (red line) at 1:150. The cells were incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of>10,000 events was performed.

Datasheet

SDS

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Datasheet Target References Q & As Review & reward Protocols Associated Products

Basic Information

Host Animal

Mouse

Clone

9B5F4

Application

ELISA, WB, IHC, IF, FC

Immunogen

Recombinant Human Programmed cell death 1 ligand 1 protein (aa 19-238).

Host Species

Mouse

Specificity

Human

Antibody Isotype

IgG2b

Clonality

Monoclonal Antibody

Application Notes

Application	Note
WB	1:1,000-1:5,000
IHC	1:50-1:200
IF	1:50-1:200

Formulations & Storage [For reference only, actual COA shall prevail!]

Format

Liquid

Buffer

PBS, Glycerol

Preservative

Sodium azide

Concentration

Batch dependent

Purity

> 95% Purity determined by SDS-PAGE.

Storage

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freezethaw cycles.

More Infomation

Target

Full Name

CD274 Molecule

Entrez Gene ID

29126

UniProt ID

Q9NZQ7

Alternative Names

CD274 Molecule; CD274 Antigen; B7 Homolog 1; Programmed Cell Death 1 Ligand 1; PDCD1 Ligand 1; PDCD1LG1; PDCD1L1;

Function

Plays a critical role in induction and maintenance of immune tolerance to self (PubMed:11015443, PubMed:28813417, PubMed:28813410).
As a ligand for the inhibitory receptor PDCD1/PD-1, modulates the activation threshold of T-cells and limits T-cell effector response (PubMed:11015443, PubMed:28813417, PubMed:28813410).
Through a yet unknown activating receptor, may costimulate T-cell subsets that predominantly produce interleukin-10 (IL10) (PubMed:10581077).
The PDCD1-mediated inhibitory pathway is exploited by tumors to attenuate anti-tumor immunity and escape destruction by the immune system, thereby facilitating tumor survival (PubMed:28813417, PubMed:28813410).
The interaction with PDCD1/PD-1 inhibits cytotoxic T lymphocytes (CTLs) effector function (By similarity).
The blockage of the PDCD1-mediated pathway results in the reversal of the exhausted T-cell phenotype and the normalization of the anti-tumor response, providing a rationale for cancer immunotherapy (By similarity).

Biological Process

Adaptive immune response Source: UniProtKB-KW
Cell surface receptor signaling pathway Source: UniProtKB
Cellular response to lipopolysaccharide Source: GO_Central
Immune response Source: UniProtKB
Negative regulation of activated T cell proliferation Source: UniProtKB
Negative regulation of CD4-positive, alpha-beta T cell proliferation Source: UniProtKB
Negative regulation of CD8-positive, alpha-beta T cell activation Source: UniProtKB
Negative regulation of interferon-gamma production Source: UniProtKB
Negative regulation of interleukin-10 production Source: UniProtKB
Negative regulation of T cell proliferation Source: GO_Central
Negative regulation of tumor necrosis factor superfamily cytokine production Source: UniProtKB
Positive regulation of activated CD8-positive, alpha-beta T cell apoptotic process Source: UniProtKB
Positive regulation of cell migration Source: Ensembl
Positive regulation of interleukin-10 production Source: UniProtKB
Positive regulation of T cell proliferation Source: GO_Central
Positive regulation of tolerance induction to tumor cell Source: UniProtKB
Response to cytokine Source: AgBase
Signal transduction Source: UniProtKB
T cell costimulation Source: UniProtKB
Toxin transport Source: Ensembl

Cellular Location

Cell membrane; Early endosome membrane; Recycling endosome membrane. Associates with CMTM6 at recycling endosomes, where it is protected from being targeted for lysosomal degradation.
Isoform 1: Cell membrane
Isoform 2: Endomembrane system

Involvement in disease

Truncation of the 3'-untranslated (3'-UTR) region of CD274 transcripts leads to elevated expression of CD274 in multiple cancers including T-cell leukemia, diffuse large B-cell lymphoma and stomach adenocarcinoma (PubMed:27281199). Disruption of 3'-UTR region is caused by structural variants that stabilize CD274 transcripts, leading to overexpression (PubMed:27281199). Increased expression in tumors promotes immune evasion and tumor cell growth by allowing malignant cells to escape destruction by the immune system (PubMed:27281199).

Topology

Extracellular: 19-238
Helical: 239-259
Cytoplasmic: 260-290

PTM

Ubiquitinated; STUB1 likely mediates polyubiquitination of PD-L1/CD274 triggering its degradation.

Verocq, C., Decaestecker, C., Rocq, L., De Clercq, S., Verrellen, A., Mekinda, Z., ... & D'haene, N. (2020). The daily practice reality of PD‑L1 (CD274) evaluation in non‑small cell lung cancer: A retrospective study. Oncology letters, 19(5), 3400-3410.

Riemann, D., Schütte, W., Turzer, S., Seliger, B., & Möller, M. (2020). High PD-L1/CD274 Expression of Monocytes and Blood Dendritic Cells Is a Risk Factor in Lung Cancer Patients Undergoing Treatment with PD1 Inhibitor Therapy. Cancers, 12(10), 2966.

Kim, J. Y., Lee, K. H., Kang, J., Borcoman, E., Saada-Bouzid, E., Kronbichler, A., ... & Gamerith, G. (2019). Hyperprogressive disease during anti-PD-1 (PDCD1)/PD-L1 (CD274) therapy: a systematic review and meta-analysis. Cancers, 11(11), 1699.

Fabrizio, F. P., Trombetta, D., Rossi, A., Sparaneo, A., Castellana, S., & Muscarella, L. A. (2018). Gene code CD274/PD-L1: from molecular basis toward cancer immunotherapy. Therapeutic advances in medical oncology, 10, 1758835918815598.

Huang, C. Y., Chiang, S. F., Ke, T. W., Chen, T. W., You, Y. S., Chen, W. T. L., & Chao, K. C. (2018). Clinical significance of programmed death 1 ligand-1 (CD274/PD-L1) and intra-tumoral CD8+ T-cell infiltration in stage II–III colorectal cancer. Scientific reports, 8(1), 1-10.

Clavé, S., Pijuan, L., Casadevall, D., Taus, Á., Gimeno, J., Hernández‐Llodrà, S., ... & Salido, M. (2018). CD 274 (PDL 1) and JAK 2 genomic amplifications in pulmonary squamous‐cell and adenocarcinoma patients. Histopathology, 72(2), 259-269.

Yang, L., Cai, Y., Zhang, D., Sun, J., Xu, C., Zhao, W., ... & Pan, C. (2018). miR-195/miR-497 regulate CD274 expression of immune regulatory ligands in triple-negative breast cancer. Journal of breast cancer, 21(4), 371-381.

Sun, J., Lian, M., Ma, H., Wang, R., Ma, Z., Wang, H., ... & Fang, J. (2018). Competing endogenous RNA network analysis of CD274, IL‑10 and FOXP3 co‑expression in laryngeal squamous cell carcinoma. Molecular medicine reports, 17(3), 3859-3869.

Lee, S. J., Jun, S. Y., Lee, I. H., Kang, B. W., Park, S. Y., Kim, H. J., ... & Kim, J. G. (2018). CD274, LAG3, and IDO1 expressions in tumor-infiltrating immune cells as prognostic biomarker for patients with MSI-high colon cancer. Journal of cancer research and clinical oncology, 144(6), 1005-1014.

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Please try the standard protocols which include: protocols, troubleshooting and guide.

Enzyme-linked Immunosorbent Assay (ELISA)
Flow Cytometry
Immunofluorescence (IF)
Immunohistochemistry (IHC)
Immunoprecipitation (IP)
Western Blot (WB)
Enzyme Linked Immunospot (ELISpot)
Proteogenomic
Other Protocols

Antibody Pairs

CD274 Matched Antibody Pair (214) (CAT#: APMAB-214LY)

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For research use only. Not intended for any clinical use.

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