Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173) is part of the multiplex kits range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.
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Key facts
- Reactive species
- Human
- Target
- CD47
(See target data below)
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Target data
Function
Adhesive protein that mediates cell-to-cell interactions (PubMed:11509594, PubMed:15383453). Acts as a receptor for thrombospondin THBS1 and as modulator of integrin signaling through the activation of heterotrimeric G proteins (PubMed:19004835, PubMed:7691831, PubMed:8550562). Involved in signal transduction, cardiovascular homeostasis, inflammation, apoptosis, angiogenesis, cellular self-renewal, and immunoregulation (PubMed:11509594, PubMed:15383453, PubMed:19004835, PubMed:27742621, PubMed:32679764, PubMed:7691831, PubMed:8550562). Plays a role in modulating pulmonary endothelin EDN1 signaling (PubMed:27742621). Modulates nitrous oxide (NO) signaling, in response to THBS1, hence playing a role as a pressor agent, supporting blood pressure (By similarity). Plays an important role in memory formation and synaptic plasticity in the hippocampus (By similarity). Receptor for SIRPA, binding to which prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells (PubMed:11509594). Interaction with SIRPG mediates cell-cell adhesion, enhances superantigen-dependent T-cell-mediated proliferation and costimulates T-cell activation (PubMed:15383453). Positively modulates FAS-dependent apoptosis in T-cells, perhaps by enhancing FAS clustering (By similarity). Plays a role in suppressing angiogenesis and may be involved in metabolic dysregulation during normal aging (PubMed:32679764). In response to THBS1, negatively modulates wound healing (By similarity). Inhibits stem cell self-renewal, in response to THBS1, probably by regulation of the stem cell transcription factors POU5F1/OCT4, SOX2, MYC/c-Myc and KLF4 (By similarity). May play a role in membrane transport and/or integrin dependent signal transduction (PubMed:7691831). May prevent premature elimination of red blood cells (By similarity).
Additional Targets
SIRPA, IDO1, LGALS9, CD276, KLRC1, CD244
Alternative names
CD47, MER6, Leukocyte surface antigen CD47, Antigenic surface determinant protein OA3, Integrin-associated protein, Protein MER6, IAP
What's included?
Recommended products
Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173) is part of the multiplex kits range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.
Key facts
- Reactive species
- Human
- Target
- CD47
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
Inhibitory Immune Checkpoint panel 2 - Human WB ab278173 contains multiple trial-sized versions of anti-human antibody clones against CD47, SIRP alpha, NKG2A, galectin 9/Gal-9, Indoleamine 2, 3-dioxygenase, 2B4 and CD276. For guidelines on how to use each antibody within the panel, please consult the individual datasheet for each antibody.specifically selected for high performance in various applications. They are provided as a sampler panel to allow you to easily evaluate each antibody.
Panel contains:
- Rabbit monoclonal [EPR21794] to CD47 (20 μL) ab218810
- Rabbit monoclonal [EPR16264] to SIRP alpha (20 μL) ab191419
- Rabbit monoclonal [EPR23620-39] to NKG2A (20 μL) ab252437
- Rabbit monoclonal [EPR22214] to galectin 9/Gal-9 (20 μL) ab227046
- Rabbit monoclonal [EPR20374] to Indoleamine 2, 3-dioxygenase (20 μL) ab211017
- Rabbit monoclonal [EPR23692-33] to 2B4 (20 μL) ab256370
- Rabbit monoclonal [SP206] to CD276 (20 μL) ab227670
Explore our range of antibody sample panels designed to provide you with a variety of trial-size antibodies in a convenient and cost-effective format.
Carrier-free formulations of our recombinant antibodies are also available for easy conjugation to labels of your choice and for multiplex applications. Use our intuitive search and select carrier-free or your label of choice. For bespoke conjugations or large volumes email bespoke@abcam.com.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
CD47 also known as integrin-associated protein (IAP) is a transmembrane protein with a mass of approximately 50 kDa. Located on the cell surface CD47 is expressed widely across various tissues. It interacts with signal regulatory protein alpha (SIRPα) which is predominantly expressed on myeloid and neural cells forming an immune checkpoint. Another significant protein NKG2A an inhibitory receptor plays a role in immune modulation. Indoleamine 23-dioxygenase (IDO) is an intracellular enzyme that catabolizes tryptophan. Meanwhile 2B4 an immune cell surface protein and Galectin-9 (Gal-9) modulate immune response. CD276 (B7-H3) acts as a co-stimulatory molecule and has a mass of roughly 80 kDa. These molecules are highly expressed in immune and tumor cells.
- Biological function summary
CD47 modulates immune evasion by binding SIRPα sending inhibitory signals to macrophages to prevent phagocytosis. The NKG2A receptor with its ligand HLA-E contributes to immune tolerance by silencing NK and T cells. IDO by degrading tryptophan facilitates immune escape in the tumor microenvironment. The protein 2B4 allows natural killer (NK) and activated T cell signaling enhancing immune surveillance. Gal-9 acts as a potent immunosuppressive factor that negatively regulates T cell activity. CD276 fosters immune cell interactions often displaying anti-tumor properties in immune cells.
- Pathways
CD47 and SIRPα interact within the phagocytosis inhibitory pathway important in regulating macrophage activity. NKG2A functions within the natural killer cell inhibitory pathway signaling through SHP-1 and SHP-2 phosphatases. IDO plays a vital role in the kynurenine pathway contributing to immune suppression. 2B4 functions in the signaling lymphocyte activation molecule (SLAM) family pathway impacting cell-mediated cytotoxicity. Gal-9 often cooperates with T cell immunoglobulin and mucin-domain containing-3 (TIM-3) to enhance immune suppression. CD276 as a member of the B7 family influences the co-stimulatory pathway regulating immune response.
- Associated diseases and disorders
CD47 associates with cancer tissues frequently contributing to tumor immune evasion. This interaction with SIRPα plays a role in preventing anti-tumor macrophage activity. NKG2A has been implicated in various cancers aiding immune escape by inhibiting NK cells. IDO contributes to immune tolerance in cancer by depleting tryptophan. 2B4 has links to autoimmune disorders where dysregulation affects immune balance. Gal-9 through its interaction with TIM-3 associates with autoimmune diseases by exacerbating immune tolerance. CD276 emerges as a potential therapeutic target in melanoma where it modulates immune responses to reduce tumor growth.
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13 product images
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-CD276 antibody [SP206] (Anti-CD276 antibody [SP206] ab227670) at 1/1000 dilution.
Lane 1: Wild-type HEK293T cell lysate, 20 ug
Lane 2: CD276 knockout HEK293T cell lysate, 20 ug
Lane 3: LNCaP cell lysate, 20 ug
Lane 4: Raji cell lysate, 20 ugSecondary (all lanes): Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 at 1/10000 dilution.
Predicted MW: 57kDa.
Observed MW: 90-110kDa
Lanes 1-4: Merged signal (red and green). Green - Anti-CD276 antibody [SP206] ab227670 observed at 90-110 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-CD276 antibody [SP206] ab227670 Anti-CD276 antibody [SP206] was shown to specifically react with CD276 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line Human CD276 knockout HEK-293T cell line ab266658 (knockout cell lysate Human CD276 knockout HEK-293T cell lysate ab257097) was used. Wild-type and CD276 knockout samples were subjected to SDS-PAGE. Anti-CD276 antibody [SP206] ab227670 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CD276 antibody [SP206] - BSA and Azide free (Anti-CD276 antibody [SP206] - BSA and Azide free ab240407) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: CD276 knockout HEK293T cell lysate at 20 µg
Lane 3: LNCaP cell lysate at 20 µg
Lane 4: Raji cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 90-110 kDa
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-CD276 antibody [SP206] (Anti-CD276 antibody [SP206] ab227670) at 1/400 dilution.Lane 1: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Predicted MW: 57 kDa.
All lanes: Western blot - Anti-CD276 antibody [SP206] - BSA and Azide free (Anti-CD276 antibody [SP206] - BSA and Azide free ab240407) at 1/400 dilution
All lanes: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Predicted band size: 57 kDa
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-2B4 antibody [EPR23692-33] (Anti-2B4 antibody [EPR23692-33] ab256370) at 1/1000 dilution.Lane 1: Human spleen tissue lysate, 20 ug
Secondary (all lanes): VeriBlot for IP Detection Reagent (HRP) ab131366 at 1/1000 dilution.
Predicted MW: 42 kDa.
Observed MW: 70 kDa
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 48 seconds.
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-2B4 antibody [EPR23692-33] (Anti-2B4 antibody [EPR23692-33] ab256370) at 1/1000 dilution.Lane 1: KG-1a (human bone marrow myeloblast) whole cell lysate, 10 ug
Secondary (all lanes): VeriBlot for IP Detection Reagent (HRP) ab131366 at 1/5000 dilution.
Predicted MW: 42 kDa.
Observed MW: 70 kDa
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 24 seconds.
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] ab211017) at 1/1000 dilution.Lane 1: Human ovary cancer lysate, 20 ug
Lane 2: Human placenta lysate, 20 ug
Lane 3: Human tonsil lysate, 20 ug
Lane 4: SK-OV-3 (Human ovarian cancer cell line) whole cell lysate, 10 ug
Secondary (all lanes): Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/100000 dilution.
Predicted MW: 45kDa.
Observed MW: 45kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-3: 3 minutes; Lane 4: 15 seconds.
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] ab211017) at 1/1000 dilution.Lane 1: Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 10 ug
Lane 2: HeLa whole cell lysate treated with 50ng/ml Interferon-gamma (IFN-gamma, Recombinant Human Interferon gamma protein ab51240) for 16 hours, 10 ug
Secondary (all lanes): Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/100000 dilution.
Predicted MW: 45kDa.
Observed MW: 45kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
The level of Indoleamine 2, 3-dioxygenase expression can be induced by IFN-γ treatment (PMID 16368976).
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-galectin 9/Gal-9 antibody [EPR22214] (Anti-galectin 9/Gal-9 antibody [EPR22214] ab227046) at 1/1000 dilution.Lane 1: THP-1 (human monocytic leukemia monocyte), whole cell lysate, 20 ug
Lane 2:HEK-293T (human embryonic kidney epithelial cell), whole cell lysate, 20 ug
Secondary (all lanes): Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/20000 dilution.
Predicted MW: 40 kDa.
Observed MW: 34-39kDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
The expression of the isoforms observed is consistent with what has been described in the literature (PMID 11886844, PMID: 22805533).
Degraded fragments (13-15 kDa) has also been described (PMID: 15811318).
Negative control: HEK-293T (PMID: 24333696).
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-galectin 9/Gal-9 antibody [EPR22214] (Anti-galectin 9/Gal-9 antibody [EPR22214] ab227046) at 1/1000 dilution.Lane 1: U937 (human histiocytic lymphoma monocyte), whole cell lysate, 20 ug
Lane 2:HL-60 (human Acute Promyelocytic Leukemia promyeloblast), whole cell lysate, 20 ug
Lane 3: MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate, 20 ug
Lane 4:PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate, 20 ugSecondary (all lanes): Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/20000 dilution.
Predicted MW: 40 kDa.
Observed MW: 34-39kDa. 13-15kDa - degradation product
Blocking/Dilution buffer: 5% NFDM/TBST.
The expression of the isoforms observed is consistent with what has been described in the literature (PMID 11886844, PMID: 22805533).
Degraded fragments (13-15 kDa) have also been descried (PMID: 15811318).
Negative control: PANC-1.
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-NKG2A antibody [EPR23620-39] (Anti-NKG2A antibody [EPR23620-39] ab252437) at 1/1000 dilution.Lane 1: HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate, 10 ug.
Lane 2: HEK-293T transfected with NKG2-A (WT) expression vector containing a myc-His-tag®, whole cell lysate, 10 ug
Lane 3: HEK-293T transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate, 10 ug
Lane 4: HEK-293T transfected with NKG2-C (WT) expression vector containing a myc-His-tag®, whole cell lysate, 10 ug
Lane 5: HEK-293T transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate, 10 ug
Lane 6: HEK-293T transfected with NKG2-E (WT) expression vector containing a myc-His-tag®, whole cell lysate, 10 ugSecondary (all lanes): Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/100000 dilution.
Predicted MW: 26 kDa.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-SIRP alpha antibody [EPR16264] (Anti-SIRP alpha antibody [EPR16264] ab191419) at 1/1000 dilution.Lane 1: Wild-type HAP1 whole cell lysate, 20 ug.
Lane 2: SIRPA knockout HAP1 whole cell lysate, 20 ug
Lane 3: THP1 whole cell lysate, 20 ug.
Lane 4: Jurkat whole cell lysate (negative control), 20 ug.Lanes 1 - 4: Merged signal (red and green). Green - Anti-SIRP alpha antibody [EPR16264] ab191419 observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
Anti-SIRP alpha antibody [EPR16264] ab191419 was shown to recognize SIRP alpha in wild-type HAP1 cells as signal was lost at the expected MW in SIRPA knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SIRPA knockout samples were subjected to SDS-PAGE. Anti-SIRP alpha antibody [EPR16264] ab191419 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-SIRP alpha antibody [EPR16264] (Anti-SIRP alpha antibody [EPR16264] ab191419) at 1/5000 dilution.Lane 1: THP1 cell lysate, 20 ug.
Lane 2: SW480 cell lysate, 20 ug
Lane 3: Human fetal brain lysate, 20 ug.
Secondary (all lanes): Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated, at 1/1000 dilution.
Predicted MW: 55kDa.
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-CD47 antibody [EPR21794] (Anti-CD47 antibody [EPR21794] ab218810) at 1/500 dilution.Lane 1: Wild-type HEK293T cell lysate, 20 ug.
Lane 2: CD47 knockout HEK293T cell lysate, 20 ug.
Lane 3: Jurkat cell lysate, 20 ug.
Lane 4:HepG2 cell lysate, 20 ug.Secondary (all lanes): Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 at 1/10000 dilution.
Predicted MW: 35 kDa.
Observed MW: 47-52kDa.
Lanes 1-4: Merged signal (red and green). Green - Anti-CD47 antibody [EPR21794] ab218810 observed at 47-52 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-CD47 antibody [EPR21794] ab218810 Anti-CD47 antibody [EPR21794] was shown to specifically react with CD47 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line Human CD47 knockout HEK-293T cell line ab266324 (knockout cell lysate Human CD47 knockout HEK-293T cell lysate ab257220) was used. Wild-type and CD47 knockout samples were subjected to SDS-PAGE. Anti-CD47 antibody [EPR21794] ab218810 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Inhibitory Immune Checkpoint panel 2 - Human WB (ab278173)
All lanes: Anti-CD47 antibody [EPR21794] (Anti-CD47 antibody [EPR21794] ab218810) at 1/5000 dilution.Lane 1: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate 20 ug.
Lane 2: U937 (human histiocytic lymphoma cell line) whole cell lysate, 20 ug.
Lane 3: A549 (human lung carcinoma cell line) whole cell lysate, 20 ug.
Lane 4: U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate, 20 ug
Lane 5: Human brain lysate, 10ug
Secondary (all lanes): Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/100000 dilution.
Predicted MW: 35 kDa.
All lanes: Anti-PD-L1 antibody [73-10] (Anti-PD-L1 antibody [73-10] ab228415) at 1/1000 dilution.
Lane 1: NCI-H1975 (human non-small cell lung cancer cell line), whole cell lysate, 20 ug.
Lane 2: Human placenta, 20 ug.
Lane 3: Human thymus, 20 ug.
Secondary (all lanes): Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/50000 dilution.
Predicted MW: 33 kDa.
Observed MW: 47-52 kDa. The molecular mass observed is consistent with the literature (PMID 12393467, PMID 11034562).
Exposure time: Lane 1: 26 seconds; Lane 2: 70 seconds; Lanes 3-4: 3 minutes; Lane 5: 3 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
We recommend that samples are not boiled after adding loading buffer as this may cause protein aggregates.
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