Stimulatory Immune Checkpoint panel 1 - Human WB (ab278176)
Overview
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Product name
Stimulatory Immune Checkpoint panel 1 - Human WB -
Species reactivity
Reacts with: Human -
Product overview
Stimulatory Immune Checkpoint panel 1 - Human WB ab278176 contains multiple trial-sized versions of anti-human antibody clones against OX40L/TNFSF4, CD134/OX40L, ICOS, ICOS Ligand/ICOSL, CD137, 4-1BBL, CD40 and TRAP/CD40L specifically selected for high performance in various applications. They are provided as a sampler panel to allow you to easily evaluate each antibody.
For guidelines on how to use each antibody within the panel, please consult the individual datasheet for each antibody.
Panel contains:
- Rabbit monoclonal [EP1168Y] to OX40L/TNFSF4 (20 μL) ab76130
- Rabbit monoclonal [EPR23000-42] to CD134 / OX40L receptor (20 μL) ab264466
- Rabbit monoclonal [EPR20560] to ICOS (20 μL) ab224644
- Rabbit monoclonal [EPR20240] to ICOS Ligand/ICOSL (20 μL) ab209262
- Rabbit monoclonal [EPR20238] to CD137 (20 μL) ab209256
- Rabbit monoclonal [EPR21995-147] to 4-1BBL (20 μL) ab254385
- Rabbit monoclonal [EPR20735] to CD40 (20 μL) ab224639
- Rabbit monoclonal [EP462E] to TRAP/CD40L (20 μL) ab52750
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Notes
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.
Directly conjugated versions of our antibodies are available and ready to use for multicolor flow cytometry or immunocytochemistry analysis. Please refer to the ‘Associated products’ section below.
Carrier-free formulations of our recombinant antibodies are also available for easy conjugation to labels of your choice and for multiplex applications. Please refer to the ‘Associated products’ section below.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 kit ab254385 - Anti-4-1BBL antibody [EPR21995-147] 2 x 10µl ab264466 - Anti-CD134 / OX40L receptor antibody [EPR23000-42] 2 x 10µl ab209256 - Anti-CD137 antibody [EPR20238] 2 x 10µl ab224639 - Anti-CD40 antibody [EPR20735] 2 x 10µl ab224644 - Anti-ICOS antibody [EPR20560] 2 x 10µl ab209262 - Anti-ICOS Ligand/ICOSL antibody [EPR20240] 2 x 10µl ab76130 - Anti-OX40L/TNFSF4 antibody [EP1168Y] 2 x 10µl ab52750 - Anti-TRAP/CD40L antibody [EP462E] 2 x 10µl -
Research areas
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Cellular localization
CD40: Secreted and Cell membrane. CD137: Membrane. ICOS: Secreted and Cell membrane. OX40L/TNFSF4: Membrane. CD134 / OX40L receptor: Membrane. ICOS Ligand/ICOSL: Membrane. 4-1BBL: Membrane. TRAP/CD40L: Secreted and Cell membrane. -
Database links
- Entrez Gene: 23308 Human
- Entrez Gene: 29851 Human
- Entrez Gene: 3604 Human
- Entrez Gene: 7292 Human
- Entrez Gene: 7293 Human
- Entrez Gene: 8744 Human
- Entrez Gene: 958 Human
- Entrez Gene: 959 Human
see all
Images
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)
All lanes: Anti-TRAP/CD40L antibody [EP462E] (ab52750) at 1/5000 dilution.
Lane 1: MOLT-4 (human lymphoblastic leukemia cell line) cell lysate, 10 ug
Secondary (all lanes): Goat anti-rabbit HRP labeled at 1/2000 dilution.
Predicted MW: 29 kDa.
Observed MW: 36 kDa.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-TRAP/CD40L antibody [EP462E] (ab52750) at 1µg/ml.
Lane 1: Human lymph node tissue lysate, 20 µg
Lane 2: Mouse lymph node tissue lysate, 20 µg
Secondary (all lanes): Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution.
Predicted MW: 29 kDa.
Observed MW: 36 kDa.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 90 seconds
Lane 2: 20 seconds
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-CD40 antibody [EPR20735] (ab224639) at 1/1000 dilution.
Lane 1: Human lymph node tissue lysate, 20 µg
Lane 2: Human lymphoma tissue lysate, 20 µg
Lane 3: Raji (human Burkitt's lymphoma cell line) whole cell lysate, 20 µgSecondary (all lanes): ab97051 at 1/20000 dilution.
Predicted MW: 30 kDa.
Observed MW: 42 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure: 15 seconds
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-4-1BBL antibody [EPR21995-147] (ab254385) at 1/1000 dilution.
Lane 1: Raji (human Burkitts lymphoma cell line) whole cell lysate, 30 µg
Lane 2: K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate, 60 µg
Secondary (all lanes): ab97051 at 1/20000 dilution.
Predicted MW: 27 kDa.
Observed MW: 26 kDa
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression control: K562 (PMID: 20670353).
Exposure time: 3 minutes.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-CD137 antibody [EPR20238] (ab209256) at 1/1000 dilution.
Lane 1: Human lymphoma lysate, 20 µg
Lane 2: Human fetal spleen lysate, 20 µg
Secondary (all lanes): ab97051 at 1/50000 dilution.
Predicted MW: 28 kDa.
Observed MW: 36 kDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
The MW observed is consistent with the literature: PMID 10528184; PMID 17560335.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-CD137 antibody [EPR20238] (ab209256) at 1/1000 dilution.
Lane 1: H9 (Human cutaneous T Lymphocyte cell line) whole cell lysate, 20 µg
Secondary (all lanes): ab97051 at 1/100000 dilution.
Predicted MW: 28 kDa.
Observed MW: 36, 40 kDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
The MW observed is consistent with the literature: PMID 10528184; PMID 17560335.
Exposure time: 3 mins.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-CD137 antibody [EPR20238] (ab209256) at 1/1000 dilution.
Lane 1: Whole cell lysate from untreated Jurkat (Human T cell leukemia cell line from peripheral blood), 10 µg
Lane 2: Whole cell lysate from Jurkat cells treated with 10ng/ml Phorbol-12-myristate-13-acetate (TPA) and 1μM Ionomycin (ab120116) for 48 hours, 10 µg
Secondary (all lanes): ab97051 at 1/100000 dilution.
Predicted MW: 28 kDa.
Observed MW: 36, 40 kDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
The expression level of CD137 can be induced by TPA and Ionomycin [PMID 14648705].
The MW observed is consistent with the literature: PMID 10528184; PMID 17560335.
Exposure time: 3 mins.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-ICOS Ligand/ICOSL antibody [EPR20240] (ab209262) at 1/20000 dilution.
Lane 1: His-tagged recombinant human ICOS ligand/ICOSL protein, 0.01 µg
Secondary (all lanes): ab97051 at 1/100000 dilution.
Predicted MW: 33 kDa.
Observed MW: 50 kDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 10 secs.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-ICOS Ligand/ICOSL antibody [EPR20240] (ab209262) at 1/1000 dilution.
Lane 1: Untreated JAR (Human placenta choriocarcinoma cell line) whole cell lysate, 20 µg
Lane 2: JAR whole cell lysate treated with PNGase F, 20 µg
Secondary (all lanes): ab97051 at 1/20000 dilution.
Predicted MW: 33 kDa.
Observed MW: 60 kDa (glycosylated form); 35 kDa (non-glycosylated form).
Blocking/Dilution buffer: 5% NFDM/TBST.
ICOS ligand/ICOSL is highly glycosylated (PMID 11983910).
Exposure time: 30 secs.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-ICOS Ligand/ICOSL antibody [EPR20240] (ab209262) at 1/1000 dilution.
Lane 1: Untreated HUVEC (Human umbilical vein endothelial cell line) whole cell lysate, 20 µg
Lane 2: HUVEC whole cell lysate treated with tumor necrosis factor-α (TNF-alpha, ab9642) 200 unit/ml for 24 hours, 20 µg
Secondary (all lanes): ab97051 at 1/20000 dilution.
Predicted MW: 33 kDa.
Observed MW: 60kDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
The level of ICOS Ligand/ICOSL expression can be elevated by TNF-alpha treatment (PMID 11983910).
Exposure time: 30 secs.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-ICOS antibody [EPR20560] (ab224644) at 1/1000 dilution.
Lane 1: Untreated human tonsil tissue lysate, 20 µg
Lane 2: Human tonsil tissue lysate treated with PNGase F, 20 µg
Secondary (all lanes): ab97051 at 1/20000 dilution.
Predicted MW: 22 kDa.
Observed MW: 22, 27-29 kDa.
Blocking and dilution buffer: 5% NFDM/TBST.
When untreated two distinct bands of 27kDa and 29kDa were observed. Treatment with PNGase F resulted in a single band with a molecular of 22kDa.
The expression profile observed is consistent with what has been described in the literature (PMID: 11169414 ).
Exposure time: 10 secs.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-ICOS antibody [EPR20560] (ab224644) at 1/1000 dilution.
Lane 1: Human tymus tissue lysate, 20 µg
Lane 2: HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate, 20 µg
Secondary (all lanes): ab97051 at 1/20000 dilution.
Predicted MW: 22 kDa.
Observed MW: 27-29 kDa.
Blocking and dilution buffer: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID: 11169414).
Exposure time: 10 secs.
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-ICOS antibody [EPR20560] (ab224644) at 1/1000 dilution.
Lane 1: His-tagged human ICOS (aa21-140-GGGGS-aa162-199) recombinant protein, 10 ng
Secondary (all lanes): ab97051 at 1/100000 dilution.
Predicted MW: 22 kDa.
Observed MW: 17 kDa.
Blocking and dilution buffer: 5% NFDM/TBST
Exposure time: 1 second
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-CD134 / OX40L receptor antibody [EPR23000-42] (ab264466) at 1/1000 dilution.
Lane 1: Human thymus tissue lysate, 10 µg
Lane 2: Human lymph node, 10 µg
Secondary (all lanes): ab131366 at 1/1000 dilution.
Predicted MW: 29 kDa.
Observed MW: 50 kDa.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
This blot was developed using a higher sensitivity ECL substrate.
The molecular weight observed is consistent with what has been described in the literature (PMID:9108415, 23897980).
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-CD134 / OX40L receptor antibody [EPR23000-42] (ab264466) at 1/1000 dilution.
Lane 1: Untreated human PBMC, whole cell lysate, 20 µg
Lane 2: Human PBMC treated with 10 μg/ml phytohaemagglutinin (PHA) for 2 days, whole cell lysate, 20 µg
Lane 3: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate, 20 µgSecondary (all lanes): ab131366 at 1/500 dilution.
Predicted MW: 29 kDa.
Observed MW: 50, 37 kDa.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 37 seconds.
The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID:9108415,23897980).
Negative control: Jurkat (PMID:9108415).
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Western blot - Stimulatory immune checkpoint panel 1 - human WB (ab278176)All lanes: Anti-OX40L/TNFSF4 antibody [EP1168Y] (ab76130) at 1/1000 dilution.
Lane 1: HUVEC (human umbilical vein endothelial cell) whole cell lysate, 20 µg
Lane 2: A-172 (human brain glioblastoma) whole cell lysate, 20 µg
Lane 3: Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate, 20 µgSecondary (all lanes): ab97051 at 1/100000 dilution.
Predicted MW: 21 kDa.
Observed MW: 30 kDa.
OX40L/TNFSF4 is a glycoprotein. The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID: 8642328)
Negative control: Raji (PMID: 8642328)
Exposure times: 6 secs.