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Rabbit Recombinant Monoclonal PD-L1 antibody. Carrier free. Suitable for ICC/IF, Flow Cyt, WB, IHC-Fr, IHC-P and reacts with Human, Recombinant full length protein - Human samples.

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Images

Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (AB209889), expandable thumbnail
  • Flow Cytometry - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (AB209889), expandable thumbnail
  • Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (AB209889), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (AB209889), expandable thumbnail
  • Immunohistochemistry (Frozen sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (AB209889), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFFlow CytWBIHC-FrIHC-P
Human
Tested
Tested
Tested
Tested
Tested
Recombinant full length protein - Human
Not recommended
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
2 µg/mL
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Recombinant full length protein - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Not recommended
Not recommended

Species
Recombinant full length protein - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Recombinant full length protein - Human
Dilution info
-
Notes

Please check the parent abID, Anti-PD-L1 antibody [28-8] ab205921, for more information on dilutions.

Anti-PD-L1 antibody [73-10] ab228415 works better than Anti-PD-L1 antibody [28-8] ab205921 in western blot testing.

Species
Human
Dilution info
-
Notes

Please check the parent abID, Anti-PD-L1 antibody [28-8] ab205921, for more information on dilutions.

Tested
Tested

Species
Human
Dilution info
1 µg/mL
Notes

-

Not recommended
Not recommended

Species
Recombinant full length protein - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
2 µg/mL
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Recombinant full length protein - Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

11 products for Alternative Version

6 products for Alternative Product

Target data

Function

Plays a critical role in induction and maintenance of immune tolerance to self (PubMed:11015443, PubMed:28813410, PubMed:28813417, PubMed:31399419). 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:28813410, PubMed:28813417, PubMed:36727298). Through a yet unknown activating receptor, may costimulate T-cell subsets that predominantly produce interleukin-10 (IL10) (PubMed:10581077). Can also act as a transcription coactivator: in response to hypoxia, translocates into the nucleus via its interaction with phosphorylated STAT3 and promotes transcription of GSDMC, leading to pyroptosis (PubMed:32929201). 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:28813410, PubMed:28813417). 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).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal PD-L1 antibody. Carrier free. Suitable for ICC/IF, Flow Cyt, WB, IHC-Fr, IHC-P and reacts with Human, Recombinant full length protein - Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
28-8
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab209889 is the Low endotoxin, azide-free version of Anti-PD-L1 antibody [28-8] ab205921.

Anti-PD-L1 antibody [28-8] has been used as detector antibody in Human PD-L1 SimpleStep ELISA® kit (ab214565).

Additional information on positive controls:

Tissue:
Tonsil- with hyperreactive changes
Note: Tonsil Specimens- is recommended to screen several hyper-reactive tonsils to find those with highest expression of PD-L1 in crypt epithelium, macrophages homing the germinal centers and interfollicular mononuclear leukocytes.

Tumor tissues- prescreened for positive tumor and inflammatory infiltrates
Note: Tumor Specimens- PD-L1 expression varies by tumor type so screening is recommended to find positive and negative tumor controls. Refer to web link publication below to find some suggested tumor types. Many tumor specimens have some inflammatory macrophages and mononuclear leukocytes. Best to look for specimens with high numbers of these cells

Cell Lines:
Positives: B-CPAP- high, ES-2- medium, HCC70 - low

For primary negative control, isotype control, RabMAb negative control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) is recommended.

For negative control sample, cell line COLO205 is recommended.

For PD-L1 protein, see Recombinant human PD-L1 protein (Active) ab167713

Recommended protocols:

For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy)

For IHC usage on FFPE tissues, the following antigen solution is recommended with clone 28-8 - Universal HIER antigen retrieval reagent (Universal HIER antigen retrieval reagent (10X) ab208572) KO Validated

For recommended Flow Cytometry (Flow Cyt) protocol, please refer to the protocol book here (downloadable copy)

Western blot usage

For clone 28-8, it is recommended to use Odyssey system. This system has the advantages of a wider dynamic range and less background than chemiluminescence.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

What does low endotoxin mean?
Our low endotoxin, azide-free formats have low endotoxin level (1 EU/mg, determined by the TAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

PD-L1 also known as Programmed Death-Ligand 1 or CD274 is a protein involved in immune modulation. Mechanically PD-L1 interacts with its receptors particularly PD-1 to regulate cellular immune responses. This transmembrane protein has a calculated molecular weight of approximately 33 kDa. PD-L1 is expressed on various cell types including tumor cells and immune cells such as dendritic cells macrophages and B cells. Its expression is often upregulated in response to inflammatory cytokines.

Biological function summary

PD-L1 plays a central role in immune evasion mechanisms utilized by tumors. It is not part of a larger protein complex but directly interacts with PD-1 and CD80. When PD-L1 binds to PD-1 it sends inhibitory signals leading to decreased T cell activation and proliferation allowing cancer cells to avoid immune destruction. PD-L1 expression provides a mechanism for tumors to suppress immune surveillance facilitating tumor progression.

Pathways

PD-L1 is integral to the immune checkpoint pathway which is an important regulator of immune response. The interaction between PD-L1 and PD-1 provides a mechanism for immune tolerance which is part of the broader adaptive immune system pathway. PD-L1 is related to other immune checkpoint proteins such as CTLA-4 in its function to limit autoreactivity and promote immune homeostasis.

Associated diseases and disorders

PD-L1 is most associated with cancer particularly in tumors such as melanoma and non-small cell lung cancer. PD-L1 expression on tumor cells often correlates with poor prognosis. PD-L1 directly interacts with PD-1 in these cancers a target for immunotherapies such as checkpoint inhibitors which aim to block this interaction and restore immune activity against tumors. PD-L1 involvement extends to autoimmune disorders where altered expression can impact tolerance and lead to immune-related tissue damage.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

24 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail
    This image is courtesy of a customer review submitted by Dr. Dimitra Kalamida.

    Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunocytochemistry/ Immunofluorescence staining of U-87 MG (human glioblastoma-astrocytoma epithelial cell line) cells using rabbit Anti-PD-L1 antibody

    Paraformaldehyde-fixed Triton X-100 permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell line) cells stained for PD-L1 (red) using Anti-PD-L1 antibody [28-8] ab205921 at 1/200 dilution in ICC/IF followed by CF568 Donkey anti-rabbit IgG(H+L) secondary antibody at 1/500 dilution.

    Alexa Fluor® 488 (Alexa Fluor® 488 Anti-PD-L1 antibody [28-8] ab209959) and Alexa Fluor® 647 (Alexa Fluor® 647 Anti-PD-L1 antibody [28-8] - Extracellular domain ab209960) conjugated versions are available for this clone.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Flow Cytometry - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Flow Cytometry - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Flow Cytometry staining of PDL-1 KO cells using rabbit Anti-PD-L1 antibody

    Anti-PD-L1 antibody [28-8] ab205921 specificity testing by Flow Cytometry (KO testing): Loss of detection on KO cells.Strong detection with anti-PD-L1 (Anti-PD-L1 antibody [28-8] ab205921 clone 28-8) TALEN constructs targeting exon4 of human PD-L1 transcript variant 1 (NM_014143.3) and complete knock out (K.O) confirmed by deep sequencing in clone L2-14. Cell surface staining is almost completely eliminated in the L2987 L2-14 KO cell line. For recommended Flow Cytometry (Flow Cyt) protocol please refer to the protocol book in the protocol section. Alexa Fluor® 488 (Alexa Fluor® 488 Anti-PD-L1 antibody [28-8] ab209959) and Alexa Fluor® 647 (Alexa Fluor® 647 Anti-PD-L1 antibody [28-8] - Extracellular domain ab209960) conjugated versions are available for this clone.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    This data was developed using Anti-PD-L1 antibody [28-8] ab205921, the same antibody clone in a different buffer formulation:

    (A and B) Western blots of recombinant PD-L1 protein (Lane 1), cell lysates of CHO-PD-L1 (Lane 3), CHO (Lane 4), ES-2 (Lane 5) and Colo205 (Lane 6) cell lines. In B, anti-PD-L1 (Anti-PD-L1 antibody [28-8] ab205921, clone 28-8) was pre-incubated with purified recombinant PDL1 protein overnight at 4°C.

    Blank/no sample (Lane2). Lane 2 is blank on purpose.

    For recommended Western Blot (WB) protocol, please refer to the protocol book in the Support&downloads section of Anti-PD-L1 antibody [28-8] ab205921

    All lanes: Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Predicted band size: 33 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human tonsil tissue using rabbit Anti-PD-L1 antibody

    IHC image of Anti-PD-L1 antibody [28-8] ab205921 staining PD-L1 in human tonsil formalin fixed paraffin embedded tissue sections* performed on a Leica BOND RX (Polymer Refine kit). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9 epitope retrieval solution 2) for 30 mins at 98°C. The section was then incubated with Anti-PD-L1 antibody [28-8] ab205921 5μg/ml working concentration for 60 mins at room temperature and detected using an HRP conjugated compact polymer system for 8 minutes at room temperature. DAB was used as the chromogen for 10 minutes at room temperature. The section was then counterstained with hematoxylin blued dehydrated cleared and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank supported by the NIHR Cambridge Biomedical Research Centre

    This image was generated using Anti-PD-L1 antibody [28-8] ab205921, the same antibody but with BSA and Azide

  • Immunohistochemistry (Frozen sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Frozen sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Frozen sections) staining of human tonsil tissue using rabbit Anti-PD-L1 antibody

    This data was developed using Anti-PD-L1 antibody [28-8] ab205921, the same antibody clone in a different buffer formulation
    IHC image of PD-L1 staining in a section of frozen normal human tonsil* performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with Anti-PD-L1 antibody [28-8] ab205921 1ugml for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.


    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions primary antibody concentration and antibody incubation times.


    *Tissue obtained from the Human Research Tissue Bank supported by the NIHR Cambridge Biomedical Research Centre

  • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunocytochemistry/ Immunofluorescence staining of CHO-PDL1 (PD-L1 stably expessed Chinese hamster ovary epithelial cell) cells using rabbit Anti-PD-L1 antibody

    This data was developed using the same antibody clone in a different buffer formulation (Anti-PD-L1 antibody [28-8] ab205921).
    Immunocytochemistry analysis of CHO-PDL1 (PD-L1 stably expessed Chinese hamster ovary epithelial cell) labeling PD-L1 with purified Anti-PD-L1 antibody [28-8] ab205921 at 1/400 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 (2 μg/ml) was used as the secondary antibody. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.32 μg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
    Negative controls: Cells not transfected with PD-L1 and both the transfected and mock tranfected cells without the primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    This IHC data was generated using the same anti-PDL1 antibody clone, 28-8, in a different buffer formulation (cat# Anti-PD-L1 antibody [28-8] ab205921).

    Immunohistochemical staining of PD-L1 in formalin fixed, paraffin embedded human non-squamous non-small cell lung cancer (NSQ-NSCLC) using Anti-PD-L1 antibody [28-8] ab205921 at a dilution of 1/400, incubated for an hour at room temperature. Heat mediated antigen retrieval was carried out in low pH buffer and the sample was blocked with peroxidase blocking buffer for 3 minutes.

    This image was courteously provided by Dr. Kai Schmitt from the Institute of Pathology, Saarbrücken-Rastpfuhl.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human placenta tissue using rabbit Anti-PD-L1 antibody

    Paraformaldehyde-fixed paraffin-embedded human placenta tissue stained for PD-L1 using Anti-PD-L1 antibody [28-8] ab205921 at 1/100 dilution in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    IHC image of Anti-PD-L1 antibody [28-8] ab205921 staining PD-L1 in PD-L1 Dynamic Range Analyte Control formalin fixed paraffin embedded cell lines (HistoCyte Laboratories), performed on a Leica BOND RX (Polymer Refine kit). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9, epitope retrieval solution 2) for 30 mins at 98°C. The section was then incubated with Anti-PD-L1 antibody [28-8] ab205921, 5μg/ml working concentration, for 60 mins at room temperature and detected using an HRP conjugated compact polymer system for 8 minutes at room temperature. DAB was used as the chromogen for 10 minutes at room temperature. The section was then counterstained with hematoxylin, blued, dehydrated, cleared and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    This image was generated using Anti-PD-L1 antibody [28-8] ab205921, the same antibody but with BSA and Azide

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Anti-PD-L1 antibody [28-8] (Anti-PD-L1 antibody [28-8] ab205921)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling PD-L1 with Anti-PD-L1 antibody [28-8] ab205921 at a dilution of 1:400. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
    This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Anti-PD-L1 antibody [28-8] ab205921 specificity testing by Immunohistochemistry (KO testing): Loss of detection on KO Cells

    Strong IHC detection with anti-PD-L1 (Anti-PD-L1 antibody [28-8] ab205921, clone 28-8) is seen in human lung adenocarcinoma tumor cell line L2987. PDL1 gene was edited in L2987 cells using TALEN constructs targeting exon4 of human PD-L1, transcript variant 1 (NM_014143.3) and complete knock out (K.O) confirmed by deep sequencing in clone L2-14. IHC detection is completely eliminated in the L2987 L2-14 K.O. cell line.

    For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book in the Support&downloads section of Anti-PD-L1 antibody [28-8] ab205921.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Immunohistochemical analysis of CHO PD-L1 cells with Anti-PD-L1 antibody [28-8] ab205921 at 2 µg/ml.
    High power view
    A) Rabbit IgG, 5 µg/mL. No staining
    B) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 1)
    C) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 3)
    D) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 4)
    E) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 5)
    F) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 6)
    G) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 7)

    All batches/lots (1,3,4,5,6,7) showed consistent results.


    Note strong, moderate, and weak (red, yellow, and white arrows respectively) plasma membrane staining of CHO PD-L1 transfected cells

    For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book in the Support&downloads section of Anti-PD-L1 antibody [28-8] ab205921.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of CHO cells using rabbit Anti-PD-L1 antibody

    Immunohistochemical analysis of CHO Parental cells with Anti-PD-L1 antibody [28-8] ab205921 at 2 μg/ml.
    High power view
    A) Rabbit IgG 5 μg/mL. No staining
    B) Anti PD-L1 2 μg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 1)
    C) Anti PD-L1 2 μg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 3)
    D) Anti PD-L1 2 μg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 4)
    E) Anti PD-L1 2 μg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 5)
    F) Anti PD-L1 2 μg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 6)
    G) Anti PD-L1 2 μg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 7)

    All batches/lots (134567) showed consistent results.

    Note absence of PD-L1 expression in CHO parental cells.

    For recommended Immunohistochemistry (IHC) protocol please refer to the protocol book in the Support&downloads section.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human lung NSCLC using rabbit Anti-PD-L1 antibody

    Immunohistochemical analysis of Human Lung NSCLC with Anti-PD-L1 antibody [28-8] ab205921 at 2 µg/ml.
    High power view
    A) Rabbit IgG, 5 µg/mL. No staining
    B) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 1)
    C) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 3)
    D) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 4)
    E) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 5)
    F) Anti PD-L1, 2 µg/mL (Anti-PD-L1 antibody [28-8] ab205921 batches 6)

    All batches/lots (1,3,4,5,6) showed consistent results.


    Note linear and complete or partial (arrows) PD-L1 staining of tumor cells. Tumor associated immune cells localized over the tumor margin exhibit positive plasma membrane staining (small arrows).

    For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book in the Support&downloads section of Anti-PD-L1 antibody [28-8] ab205921.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human non-small cell lung cancer tissue labelling PD-L1 with Anti-PD-L1 antibody [28-8] ab205921. Tumor cells and immuno cells localized within the stroma show PD-LA plasma membrane staining.

    For antigen retrival buffer, Universal HIER antigen retrieval reagent (Universal HIER antigen retrieval reagent (10X) ab208572) was used.
    For IHC detection kit, Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) is recommended.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human melanoma tissue using rabbit Anti-PD-L1 antibody

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue labelling PD-L1 with Anti-PD-L1 antibody [28-8] ab205921. Tumor cells show weak and partial postive PD-L1 expresseion in the plasma membrane. PD-L1 positive tumor associated immunoe cells are also stained.

    For antigen retrival buffer Universal HIER antigen retrieval reagent (Universal HIER antigen retrieval reagent (10X) ab208572) was used.
    For IHC detection kit Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) is recommended.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human head and neck squamous cell carcinoma tissue labeling PD-L1 with Anti-PD-L1 antibody [28-8] ab205921 at 2 µg/ml. Counterstained with Hematoxylin.

    For antigen retrival buffer, Universal HIER antigen retrieval reagent (Universal HIER antigen retrieval reagent (10X) ab208572) was used.
    For IHC detection kit, Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) is recommended.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of L2987 cells using rabbit Anti-PD-L1 antibody

    Immunohistochemical analysis of formalin-fixed paraffin-embedded L2987 (Human lung adenocarcinoma cell line with endogenous PD-L1 expression) cells labeling PD-L1 with Anti-PD-L1 antibody [28-8] ab205921 at 2 μg/ml. Counterstained with Hematoxylin.

    For antigen retrival buffer Universal HIER antigen retrieval reagent (Universal HIER antigen retrieval reagent (10X) ab208572) was used.
    For IHC detection kit Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) is recommended.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PD-L1 antibody [28-8] ab205921).

  • Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

    All lanes: Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    All lanes: NCI-H1975 (human non-small cell lung cancer) whole cell lysate at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)

    Predicted band size: 33 kDa

    Observed band size: 40-60 kDa

    Exposure time: 3min

  • Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

    All lanes: Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Lane 1: CHO-S cell lysate at 15 µg

    Lane 2: Human PD-L1 transfected CHO-S cell lysate at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)

    Predicted band size: 33 kDa

    Observed band size: 40-60 kDa

    Exposure time: 1min

  • Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Western blot - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Anti-PD-L1 antibody [73-10] ab228415 works better than Anti-PD-L1 antibody [28-8] ab205921 in western blot testing.
    Blocking/Diluting buffer and concentration: 5% NFDM/TBST
    This data was developed using Anti-PD-L1 antibody [28-8] ab205921, the same antibody clone in a different buffer formulation.

    All lanes: Western blot - Anti-PD-L1 antibody [28-8] (Anti-PD-L1 antibody [28-8] ab205921) at 1/1000 dilution

    Lane 1: MDA-MB-231(human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: U-87 MG (human glioblastoma astrocytoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

    Lane 5: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 6: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 40-60 kDa

    Exposure time: 100s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human tonsil using rabbit Anti-PD-L1 antibody

    Immunohistochemical analysis of formalin-fixed paraffin-embedded human tonsil labelling PD-L1 with Anti-PD-L1 antibody [28-8] ab205921 at a dilution of 2µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). Anti-PD-L1 antibody [28-8] ab205921 anti PD-L1 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

    This data was developed using Anti-PD-L1 antibody [28-8] ab205921, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    PD-L1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human placenta using rabbit Anti-PD-L1 antibody

    Immunohistochemical analysis of formalin-fixed paraffin-embedded human placenta labelling PD-L1 with Anti-PD-L1 antibody [28-8] ab205921 at a dilution of 2µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). Anti-PD-L1 antibody [28-8] ab205921 anti PD-L1 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

    This data was developed using Anti-PD-L1 antibody [28-8] ab205921, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Tissue Microarrays stained for Anti-PD-L1 antibody [28-8] using Anti-PD-L1 antibody [28-8] ab205921 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Universal HIER antigen retrieval reagent (10X) ab208572 (Universal HIER antigen retrieval reagent). The sections were incubated with Anti-PD-L1 antibody [28-8] ab205921 at +4°C overnight. For IHC detection kit, Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) is recommended.

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