Invitrogen

CD274 (PD-L1, B7-H1) Monoclonal Antibody (MIH1), Alexa Fluor™ 700, eBioscience™

Product Image Gallery

CD274 (PD-L1, B7-H1) Antibody in Flow Cytometry (Flow) — Normal human peripheral blood cells were unstimulated (left) or stimulated with PHA-L overnight (right). Cells were then stained with Mouse IgG1 kappa Isotype Control, Alexa Fluor 700 (Product # 56-4714-81) (blue histogram) or CD274 Monoclonal Antibody, Alexa Fluor 700 (purple histogram). Cells in the lymphocyte gate were used for analysis.

CD274 (PD-L1, B7-H1) Antibody in Immunocytochemistry (ICC/IF) — Fig. 4 NEK2 positively correlates and interacts with PD-L1 in pancreatic cancer. a Western blot analysis of NEK2 and PD-L1 in clinical pancreatic tissue samples from patients ( n = 20) (N: Normal pancreatic tissue; T: Pancreatic tumor tissue). b - d Representative images and statistical results of IHC staining of NEK2 and PD-L1 in a tissue microarray ( n = 156). e - g Cell lysates from SW1990, KPC, and CFPAC-1 separately analyzed by IP and Western blotting using the antibodies indicated. Representative image is shown n = 3 independent experiments. h GST-pull down assay of NEK2-His and GST-PD-L1 protein. Representative image is shown n = 3 independent experiments. i Representative images of individual immunofluorescence staining of NEK2 and PD-L1 interaction in KPC cells by Duolink assay. The red dots (NEK2/PD-L1 interaction) indicate their interaction. Representative image is shown n = 3 independent experiments. The Spearman correlations and p -values by Spearman's test are indicated in d .

CD274 (PD-L1, B7-H1) Antibody in Neutralization (Neu) — Figure 3 TCR + PD-1 + 2D3 cells as a model assay for evaluation of involvement of PD-1 signaling in cell-mediated antigen-specific T-cell activation ( A - C ) WT1 (A, B) and gp100 (A, C) specific T-cell activation expressed as percentage viable CD8 + eGFP + PD-1 - and PD-1 + 2D3 cells (+- SEM) after 24 hours co-culture with different PD-L1 + stimulator cells. Neutralizing antibody against PD-1 (alphaPD-1; A, B; 15 ug/mL in WT1 model (A, B), 5 ug/mL in gp100 model (A)) or PD-L1 (alphaPD-L1; C) was added to cells 1 hour prior to co-culture to verify PD-1-mediated signaling, where indicated. (A) PD-1-dependent stimulating capacity of two differently generated peptide-pulsed mature monocyte-derived dendritic cells (WT1 (4 DC donors tested in two independent experiments); gp100 (4 DC donors in four independent experiments)). (B)Impact of induced PD-L1 expression on peptide-pulsed THP-1 leukemic cells on WT1-specific T-cell activation (6 replicate experiments). (C) gp100-specific PD-1 + T cell-activating capacity of peptide-pulsed wild-type or stably transduced PD-L1 + MCF-7 breast carcinoma cells (4 replicate experiments). Data information: in A, the horizontal line represents the median percentage eGFP expression ( n = 4). * P <= 0.05, ** P < 0.01, *** P < 0.001 (repeated measures one-way ANOVA with Bonferroni post-hoc test). Abbreviations: eGFP, enhanced green fluorescent protein; gp100, glycoprotein 100; PD-1, programmed death 1 protein; PD-L1, programmed death-ligand 1; TCR, T

Product Details

56-5983-42

Applications	Tested Dilution	Publications
Immunocytochemistry (ICC/IF)	-	View 2 publications 2 publications
Flow Cytometry (Flow)	5 µL (1.0 µg)/test	View 7 publications 7 publications
Neutralization (Neu)	-	View 2 publications 2 publications

Product Specifications
Species Reactivity	Human
Published species	Not Applicable
Host/Isotype	Mouse / IgG1, kappa
Recommended Isotype Control	Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), Alexa Fluor™ 700, eBioscience™
Class	Monoclonal
Type	Antibody
Clone	MIH1
Conjugate	Alexa Fluor™ 700 Alexa Fluor 488 APC APC-eFluor 780 Biotin eFluor 450 Functional Grade PE PE-Cyanine5 PE-Cyanine7 PE-eFluor 610 PerCP-eFluor 710 Unconjugated
Excitation/Emission Profile	View spectra
Form	Liquid
Concentration	5 µL/Test
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2, with 0.2% BSA
Contains	0.09% sodium azide
Storage conditions	4° C, store in dark, DO NOT FREEZE!
RRID	AB_2802351

Product Specific Information

Description: The MIH1 monoclonal antibody reacts with human B7-H1, also known as PD-L1. B7-H1, a member of the B7 family, has a predicted molecular weight of approximately 40 kDa and belongs to the Ig superfamily. B7-H1 is expressed on a majority of leukocytes. B7-H1 is a ligand for PD-1. Interaction of PD-1 with either PD-L1 (B7-H1) or PD-L2 (B7-DC) results in inhibition of T and B cell responses. MIH1 is reported to be a blocking antibody.

Applications Reported: This MIH1 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This MIH1 antibody has been pre-diluted and tested by flow cytometric analysis of stimulated normal human peripheral blood cells. This may be used at 5 µL (1.0 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Alexa Fluor 700 emits at 723 nm and can be excited with the red laser (633-647 nm). Most instruments will require a 685 LP mirror and 710/20 band pass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

Excitation: 633-647 nm; Emission: 723 nm; Laser: Red Laser

Target Information

Programmed death receptor ligand 1 (PD-L1, also called B7-H1) is a recently described B7 family member. To date, one specific receptor has been identified that can be ligated by PD-L1. This receptor, programmed death receptor 1 (PD-1), has been shown to negatively regulate T-cell receptor (TCR) signaling. Upon ligating its receptor, PD-L1 has been reported to decrease TCR-mediated proliferation and cytokine production. PD-L1 expression was found to be abundant on many murine and human cancers and could be further up-regulated upon IFN-gamma stimulation. Thus, PD-L1 might play an important role in tumor immune evasion.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Product References

NEK2 inhibition triggers anti-pancreatic cancer immunity by targeting PD-L1.

Nature communications

Zhang X,Huang X,Xu J,Li E,Lao M,Tang T,Zhang G,Guo C,Zhang X,Chen W,Yadav DK,Bai X,Liang T

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Immunocytochemistry

Tue Jul 27 00:00:00 EDT 2021

Protocol for high-throughput compound screening using flow cytometry in THP-1 cells.

STAR protocols

Spangenberg SH,Zavareh RB,Lairson LL

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Flow Cytometry

Fri Jun 18 00:00:00 EDT 2021

Protein kinase D3 regulates the expression of the immunosuppressive protein, PD‑L1, through STAT1/STAT3 signaling.

International journal of oncology

Cui B,Chen J,Luo M,Wang L,Chen H,Kang Y,Wang J,Zhou X,Feng Y,Zhang P

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Flow Cytometry

Wed Apr 01 00:00:00 EDT 2020

Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms.

Journal for immunotherapy of cancer

Neuwelt AJ,Kimball AK,Johnson AM,Arnold BW,Bullock BL,Kaspar RE,Kleczko EK,Kwak JW,Wu MH,Heasley LE,Doebele RC,Li HY,Nemenoff RA,Clambey ET

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Flow Cytometry

Wed Apr 01 00:00:00 EDT 2020

Quantitative Interactomics in Primary T Cells Provides a Rationale for Concomitant PD-1 and BTLA Coinhibitor Blockade in Cancer Immunotherapy.

Cell reports

Celis-Gutierrez J,Blattmann P,Zhai Y,Jarmuzynski N,Ruminski K,Grégoire C,Ounoughene Y,Fiore F,Aebersold R,Roncagalli R,Gstaiger M,Malissen B

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Flow Cytometry

Tue Jun 11 00:00:00 EDT 2019

Phospho‑STAT1 expression as a potential biomarker for anti‑PD‑1/anti‑PD‑L1 immunotherapy for breast cancer.

International journal of oncology

Nakayama Y,Mimura K,Tamaki T,Shiraishi K,Kua LF,Koh V,Ohmori M,Kimura A,Inoue S,Okayama H,Suzuki Y,Nakazawa T,Ichikawa D,Kono K

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Flow Cytometry

Sat Jun 01 00:00:00 EDT 2019

Generation of Tumor-Reactive T Cells by Co-culture of Peripheral Blood Lymphocytes and Tumor Organoids.

Cell

Dijkstra KK,Cattaneo CM,Weeber F,Chalabi M,van de Haar J,Fanchi LF,Slagter M,van der Velden DL,Kaing S,Kelderman S,van Rooij N,van Leerdam ME,Depla A,Smit EF,Hartemink KJ,de Groot R,Wolkers MC,Sachs N,Snaebjornsson P,Monkhorst K,Haanen J,Clevers H,Schumacher TN,Voest EE

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Flow Cytometry

Thu Sep 06 00:00:00 EDT 2018

A versatile T cell-based assay to assess therapeutic antigen-specific PD-1-targeted approaches.

Oncotarget

Versteven M,Van den Bergh JMJ,Broos K,Fujiki F,Campillo-Davo D,De Reu H,Morimoto S,Lecocq Q,Keyaerts M,Berneman Z,Sugiyama H,Van Tendeloo VFI,Breckpot K,Lion E

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Neutralization

Tue Jun 12 00:00:00 EDT 2018

PLoS neglected tropical diseases

Narasimhan PB,Akabas L,Tariq S,Huda N,Bennuru S,Sabzevari H,Hofmeister R,Nutman TB,Tolouei Semnani R

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Flow Cytometry

Sun Apr 01 00:00:00 EDT 2018

Hormonal vitamin D up-regulates tissue-specific PD-L1 and PD-L2 surface glycoprotein expression in humans but not mice.

The Journal of biological chemistry

Dimitrov V,Bouttier M,Boukhaled G,Salehi-Tabar R,Avramescu RG,Memari B,Hasaj B,Lukacs GL,Krawczyk CM,White JH

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Immunocytochemistry

Fri Dec 15 00:00:00 EST 2017

Upregulation of Programmed Death-1 and Its Ligand in Cardiac Injury Models: Interaction with GADD153.

PloS one

Baban B,Liu JY,Qin X,Weintraub NL,Mozaffari MS

Published figure using CD274 (PD-L1, B7-H1) monoclonal antibody (Product # 56-5983-42) in Neutralization

Tue Apr 12 00:00:00 EDT 2016

Bioinformatics

Protein Aliases: B7 homolog 1; B7-H1; CD274; CD274 antigen; PD-L1; PDCD1 ligand 1; pdl-1; Programmed cell death 1 ligand 1; sCD274; soluble CD274; soluble PD L1; sPD L1

Gene Aliases: B7-H; B7H1; CD274; PD-L1; PDCD1L1; PDCD1LG1; PDL1

UniProt ID: (Human) Q9NZQ7

Entrez Gene ID: (Human) 29126

Molecular Function: immunoglobulin receptor superfamily