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PD-L1 Recombinant Rabbit Monoclonal Antibody [PSH04-80]

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Catalog# HA722184

PD-L1 Recombinant Rabbit Monoclonal Antibody [PSH04-80]

Application

  • WB

  • IHC-P

  • mIHC

Reactivity

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

PD-L1 Recombinant Rabbit Monoclonal Antibody [PSH04-80]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse PD-L1 aa 1-250 / 290.

Species Reactivity

Mouse

Validated Applications

WB, IHC-P, mIHC

Molecular Weight

Predicted band size: 33 kDa

Positive Control

RAW264.7 treated with 10μg/mL LPS for 8 hours cell lysate, J774A.1 treated with 1μg/mL LPS for 24 hours cell lysate, mouse spleen tissue lysate, mouse thymus tissue, mouse lung tissue.

Conjugation

unconjugated

Clone Number

PSH04-80

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:200

  • mIHC

  • 1:500

Target

Function

Programmed death-ligand 1 (PD-L1) also known as cluster of differentiation 274 (CD274) or B7 homolog 1 (B7-H1) is a protein that in humans is encoded by the CD274 gene. Programmed death-ligand 1 (PD-L1) is a 40kDa type 1 transmembrane protein that has been speculated to play a major role in suppressing the adaptive arm of immune systems during particular events such as pregnancy, tissue allografts, autoimmune disease and other disease states such as hepatitis. Normally the adaptive immune system reacts to antigens that are associated with immune system activation by exogenous or endogenous danger signals. In turn, clonal expansion of antigen-specific CD8+ T cells and/or CD4+ helper cells is propagated. The binding of PD-L1 to the inhibitory checkpoint molecule PD-1 transmits an inhibitory signal based on interaction with phosphatases (SHP-1 or SHP-2) via Immunoreceptor Tyrosine-Based Switch Motif (ITSM). This reduces the proliferation of antigen-specific T-cells in lymph nodes, while simultaneously reducing apoptosis in regulatory T cells (anti-inflammatory, suppressive T cells) – further mediated by a lower regulation of the gene Bcl-2.

Background References

1. Lei Q et al. Resistance Mechanisms of Anti-PD1/PDL1 Therapy in Solid Tumors. Front Cell Dev Biol. 2020 Jul

2. Tran-Nguyen VK et al. Structure-based virtual screening for PDL1 dimerizers: Evaluating generic scoring functions. Curr Res Struct Biol. 2022 Jun

Subcellular Location

Cell membrane, Early endosome membrane, Recycling endosome membrane.

UNIPROT

SwissProt: Q9EP73 Mouse

Synonyms

B7 H antibody

B7 H1 antibody

B7 homolog 1 antibody

B7-H1 antibody

B7H antibody

B7H1 antibody

CD 274 antibody

CD-274 antibody

CD274 antibody

CD274 antigen antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of PD-L1 on different lysates with Rabbit anti-PD-L1 antibody (<a href="/products/HA722184" style="font-weight: bold;text-decoration: underline;">HA722184</a>) at 1/2,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: RAW264.7 treated with 10μg/mL LPS for 8 hours cell lysate<br />Lane 3: J774A.1 cell lysate<br />Lane 4: J774A.1 treated with 1μg/mL LPS for 24 hours cell lysate<br />Lane 5: Mouse spleen tissue lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 45-60 kDa<br /><br />Exposure time: 46 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722184" style="font-weight: bold;text-decoration: underline;">HA722184</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of PD-L1 on different lysates with Rabbit anti-PD-L1 antibody (HA722184) at 1/2,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: RAW264.7 treated with 10μg/mL LPS for 8 hours cell lysate
    Lane 3: J774A.1 cell lysate
    Lane 4: J774A.1 treated with 1μg/mL LPS for 24 hours cell lysate
    Lane 5: Mouse spleen tissue lysate

    Lysates/proteins at 30 µg/Lane.

    Predicted band size: 33 kDa
    Observed band size: 45-60 kDa

    Exposure time: 46 seconds; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722184) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Fluorescence multiplex immunohistochemical analysis of mouse spleen (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-PD-L1 (<a href="/products/HA722184" style="font-weight: bold;text-decoration: underline;">HA722184</a>, Green) and anti-Ki67 (<a href="/products/HA721115" style="font-weight: bold;text-decoration: underline;">HA721115</a>, Violet) on spleen. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit&trade;MH010101, www.luminiris.cn). The section was incubated in two rounds of staining: in the order of <a href="/products/HA722184" style="font-weight: bold;text-decoration: underline;">HA722184</a> (1/500 dilution) and <a href="/products/HA721115" style="font-weight: bold;text-decoration: underline;">HA721115</a> (1/2,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

    Fluorescence multiplex immunohistochemical analysis of mouse spleen (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-PD-L1 (HA722184, Green) and anti-Ki67 (HA721115, Violet) on spleen. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in two rounds of staining: in the order of HA722184 (1/500 dilution) and HA721115 (1/2,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

  • Immunohistochemical analysis of paraffin-embedded mouse thymus tissue with Rabbit anti-PD-L1 antibody (<a href="/products/HA722184" style="font-weight: bold;text-decoration: underline;">HA722184</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722184" style="font-weight: bold;text-decoration: underline;">HA722184</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse thymus tissue with Rabbit anti-PD-L1 antibody (HA722184) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722184) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-PD-L1 antibody (<a href="/products/HA722184" style="font-weight: bold;text-decoration: underline;">HA722184</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722184" style="font-weight: bold;text-decoration: underline;">HA722184</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-PD-L1 antibody (HA722184) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722184) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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Citation