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Rabbit Recombinant Monoclonal PD-L1 antibody. Carrier free. Suitable for mIHC, IHC-P, Flow Cyt, WB, ICC/IF and reacts with Human, Transfected cell line samples.


Images

Flow Cytometry - Anti-PD-L1 antibody [28-8] - BSA and Azide free (AB228413), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (AB228413), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (AB228413), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (AB228413), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (AB228413), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% 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
mIHCIHC-PFlow CytWBICC/IF
Human
Tested
Tested
Tested
Tested
Expected
Transfected cell line
Not recommended
Tested
Not recommended
Not recommended
Tested

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Transfected cell line

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

For antigen buffer for FFPE tissue, it is recommended to use Universal HIER antigen retrieval reagent (Universal HIER antigen retrieval reagent (10X) ab208572).

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

Species

Transfected cell line

Dilution info

-

Notes

For antigen buffer for FFPE tissue, it is recommended to use Universal HIER antigen retrieval reagent (Universal HIER antigen retrieval reagent (10X) ab208572).

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

Tested
Tested

Species

Human

Dilution info

-

Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Not recommended
Not recommended

Species

Transfected cell line

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

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

Not recommended
Not recommended

Species

Transfected cell line

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell line

Dilution info

-

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

10 products for Alternative Version

5 products for Alternative Product

Target data

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).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal PD-L1 antibody. Carrier free. Suitable for mIHC, IHC-P, Flow Cyt, WB, ICC/IF and reacts with Human, Transfected cell line samples.

Alternative names

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 long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab228413 is the carrier-free version of abab205921.

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. Look for specimens with high numbers of these cells
Cell Lines: Positives: B-CPAP- high, ES-2- medium, HCC70 - low

For IHC on FFPE tissues, antigen retrieval buffer (Universal HIER antigen retrieval reagent (10X) ab208572) and IHC detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) is recommended.

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

For negative control sample, use cell line COLO205, see Anti-Glypican 3 antibody [SP86] ab95363.
For PD-L1 protein, see Recombinant human PD-L1 protein (Active) ab167713

Recommended protocols:
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)

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.

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

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

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

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.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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.

16 product images

  • Flow Cytometry - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413), expandable thumbnail

    Flow Cytometry - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    PD-L1 immunohistochemistry staining of human tonsil 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 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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    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.

    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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    PD-L1 immunohistochemistry staining of human placenta 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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    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 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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    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.

    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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    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.

    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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    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.

    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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    PD-L1 immunohistochemistry 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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    PD-L1 immunohistochemistry staining of 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 (13456) 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.

    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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    PD-L1 immunohistochemistry 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).

  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413), expandable thumbnail

    Multiplex immunohistochemistry - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

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

    Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue staining PD-L1 with Anti-PD-L1 antibody [28-8] ab205921 at a 1:500 (2.19 ug/ml) dilution; PD1 with Anti-PD1 antibody [CAL20] ab237728 at 1:2000 (0.525 ug/ml) dilution and CD68 with Anti-CD68 antibody [EPR20545] ab213363 at 1:500 (1.26 ug/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

    Panel A: merged staining of anti-CD68 (magenta; Opal690), anti-PD-L1 (green; Opal520) and anti-PD1 (red; Opal570) on human tonsil.

    Panel B: anti-PD-L1 stained on cells involved in T cell inhibition.

    Panel C: anti-PD1 stained on antigen-stimulated T cells.

    Panel D: anti-CD68 stained on macrophages.

    The section was incubated in three rounds of staining: in the order of Anti-CD68 antibody [EPR20545] ab213363 and Anti-PD-L1 antibody [28-8] ab205921 for 30 mins, then Anti-PD1 antibody [CAL20] ab237728 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    PD-L1 immunohistochemistry 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] - BSA and Azide free (ab228413), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    PD-L1 immunohistochemistry 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.

  • Western blot - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413), expandable thumbnail

    Western blot - Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)

    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

    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

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