PD-L1 Antibody | Affinity Biosciences

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Western blot analysis of extracts from various samples, using CD274 Antibody.

Fig.

Figure 8| qRT-PCR and western blot results.

Fig 7.

Figure2 qRT-PCR and Western blot results (a) show that CD274 and IL8 were upregulated in the basal-like breast cancer cell lines, BT549 and MDA-MB-231 (p.

Figure 3.

Fig.

FIGURE 2 Association of JAK2/STAT3 signaling pathway activation with acquired radioresistance.

Product:	PD-L1 Antibody
Catalog:	DF6526
Description:	Rabbit polyclonal antibody to PD-L1
Application:	WB
Reactivity:	Human, Mouse, Rat
Prediction:	Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.:	33kDa, 40~70kD(Glycosylated); 33kD(Calculated).
Uniprot:	Q9NZQ7
RRID:	AB_2838488

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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MS Report

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Data Sheet

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Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Prediction:

Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(92%)

Clonality:

Polyclonal

Specificity:

CD274 Antibody detects endogenous levels of total CD274.

RRID:

AB_2838488
Cite Format: Affinity Biosciences Cat# DF6526, RRID:AB_2838488.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

B7 H; B7 H1; B7 homolog 1; B7-H1; B7H; B7H1; CD 274; CD274; CD274 antigen; CD274 molecule; MGC142294; MGC142296; OTTHUMP00000021029; PD L1; PD-L1; PD1L1_HUMAN; PDCD1 ligand 1; PDCD1L1; PDCD1LG1; PDL 1; PDL1; Programmed cell death 1 ligand 1; Programmed death ligand 1; RGD1566211;

Immunogens

Immunogen:

Uniprot:

Q9NZQ7(PD1L1_HUMAN) >>Visit HPA database.

Gene(ID):

CD274(29126)

Expression:

Q9NZQ7 PD1L1_HUMAN:

Highly expressed in the heart, skeletal muscle, placenta and lung. Weakly expressed in the thymus, spleen, kidney and liver. Expressed on activated T- and B-cells, dendritic cells, keratinocytes and monocytes.

Description:

Programmed cell death ligand 1(CD274, or B7-H1, PD-L1), is the first member of B7 family to be discovered. B7 family molecules are type I transmembrane proteins belonging to the immunoglobulin superfamily. In concert with their CD28 family receptors, the B7s are key regulators of the adaptive immune response. CD274 is suggested a negative regulator of T and B cell, and play important role in mediating tolerance of lymphocytes to self-antigens. It also involved in the costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNG, in an IL2-dependent and a PDCD1-independent manner.

Sequence:

Q9NZQ7 PD1L1_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Horse

100

Bovine

100

Sheep

100

Rabbit

100

Dog

Pig

Xenopus

Zebrafish

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9NZQ7 As Substrate

Q9NZQ7

Site	PTM Type	Source
N192	N-Glycosylation	Uniprot
S195	Phosphorylation	Uniprot
K270	Acetylation	Uniprot
S283	Phosphorylation	Uniprot
T290	Phosphorylation	Uniprot

Research Backgrounds

Q9NZQ7_PD1L1_HUMAN

Function:

Plays a critical role in induction and maintenance of immune tolerance to self. As a ligand for the inhibitory receptor PDCD1/PD-1, modulates the activation threshold of T-cells and limits T-cell effector response. Through a yet unknown activating receptor, may costimulate T-cell subsets that predominantly produce interleukin-10 (IL10).

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. 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).

PTMs:

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

Subcellular Location:

Cell membrane>Single-pass type I membrane protein. Early endosome membrane>Single-pass type I membrane protein. Recycling endosome membrane>Single-pass type I membrane protein.
Note: Associates with CMTM6 at recycling endosomes, where it is protected from being targeted for lysosomal degradation.

Cell membrane>Single-pass type I membrane protein.

Endomembrane system>Single-pass type I membrane protein.

Tissue Specificity:

Subunit Structure:

Interacts with PDCD1. Interacts (via transmembrane domain) with CMTM4 and CMTM6.

Family&Domains:

Belongs to the immunoglobulin superfamily. BTN/MOG family.

Research Fields

· Environmental Information Processing > Signaling molecules and interaction > Cell adhesion molecules (CAMs). (View pathway)

References

1). A Hybrid Nanoadjuvant Simultaneously Depresses PD-L1/TGF-β1 and Activates cGAS-STING Pathway to Overcome Radio-Immunotherapy Resistance. Advanced materials (Deerfield Beach, Fla.), 2024 (PubMed: 38229577) [IF=29.4]

2). Mitochondrial Disruption Nanosystem Simultaneously Depressed Programmed Death Ligand-1 and Transforming Growth Factor-β to Overcome Photodynamic Immunotherapy Resistance. ACS nano, 2024 (PubMed: 38227812) [IF=17.1]

3). Mitochondrial metabolism blockade nanoadjuvant reversed immune-resistance microenvironment to sensitize albumin-bound paclitaxel-based chemo-immunotherapy. Acta pharmaceutica Sinica. B, 2024 (PubMed: 39309498) [IF=14.5]

4). LncRNA MIAT correlates with immune infiltrates and drug reactions in hepatocellular carcinoma. International Immunopharmacology, 2020 (PubMed: 33221703) [IF=5.6]

Application: WB Species: Human Sample: hepatocellular carcinoma tissue

Fig 7. The relationship between MIAT and PD-L1 and sorafenib resistance was verified in vitro. (A) There was a positive correlation between MIAT and PD-L1 expression in HCC tissue samples. (B) The expression of MIAT was positively correlated with CTLA4 in HCC tissue samples. (C) In HCC cells, the expression of PD-L1 mRNA decreased after the knockdown of MIAT. (D) Knockdown of MIAT down regulated the expression of PD-L1 protein in HCC cells. (E) The level of MIAT increased after sorafenib treatment. (F) The level of PD-L1 increased after sorafenib treatment. (G) MIAT affects the proliferation of HCC cells. *: P < 0.05; **: P < 0.01; ***: P < 0.001; ****: P < 0.0001.

5). miR‑576‑3p overexpression enhances cisplatin sensitivity of ovarian cancer cells by dysregulating PD‑L1 and cyclin D1. Molecular Medicine Reports, 2021 (PubMed: 33236151) [IF=3.4]

6). Homeobox D9 drives the malignant phenotypes and enhances the Programmed death ligand-1 expression in non-small cell lung cancer cells via binding to Angiopoietin-2 promoter. World Journal of Surgical Oncology, 2023 (PubMed: 36907878) [IF=3.2]

Application: WB Species: Human Sample: NSCLC cells

Fig. 7 HOXD9 has the ability to stimulate PD-L1 expression in NSCLC cells. NCI-H661 cells were transfected with exHOXD9 or empty vector, while NCI-H292 cells were transfected with siHOXD9#1, siHOXD9#2 or their negative control siNC. A After 48 h of transfection, total PD-L1 protein expression in two NSCLC cells (NCI-H661 and NCI-H292) was evaluated by western blot assay. B After 48 h of transfection, soluble PD-L1 protein expression in two NSCLC cells was evaluated by ELISA assay. C After 48 h of transfection, membrane PD-L1 protein expression in NCI-H292 cells was evaluated by flow cytometry analysis. MFI, relative mean fluorescence intensity. ** p

7). lncRNA MIAT targets miR‐411‐5p/STAT3/PD‐L1 axis mediating hepatocellular carcinoma immune response. INTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, 2022 (PubMed: 35429078) [IF=3.0]

8). Baicalein Enhances Radiosensitivity in Colorectal Cancer via JAK2/STAT3 Pathway Inhibition. Chemical biology & drug design, 2024 (PubMed: 39152534) [IF=3.0]

Application: WB Species: Mouse Sample: CT26-R Cells

FIGURE 2 Association of JAK2/STAT3 signaling pathway activation with acquired radioresistance. Comparison of the expression levels of p-STAT3 (Tyr705), JAK2, PD-L1, and SOCS3 between parental (CT26) and acquired radioresistance (CT26-R) cells. *p

9). PA-MSHA Regulates PD-L1 Expression in Hepatoma Cells. Immunological Investigations, 2023 (PubMed: 36762677) [IF=2.8]

10). Screening of immunosuppressive factors for biomarkers of breast cancer malignancy phenotypes and subtype-specific targeted therapy. PeerJ, 2022 (PubMed: 31293831) [IF=2.7]

Application: WB Species: human Sample: T549 and MDA-MB-231 cell； MCF7 and T47D cell

Figure 8| qRT-PCR and western blot results. (A, B) qRT-PCR results showed that CD274 and IL8 were upregulated in the basal-like breast cancer cell lines BT549 and MDA-MB-231 (p < 0.0001). Similar to the qRT-PCR results, the western blot analysis (C–E) indicated that CD274 and IL8 protein were increased in the BT549 and MDA-MB-231 cell lines compared to the MCF7 and T47D cell lines. P < 0.001, p < 0.01, and p < 0.05

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PD-L1 Antibody - #DF6526