重组Anti-Granzyme B抗体[EPR22645-206] (ab255598)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22645-206] to Granzyme B
- Suitable for: WB, IHC-P, IHC-Fr, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Granzyme B抗体[EPR22645-206]
参阅全部 Granzyme B 一抗 -
描述
兔单克隆抗体[EPR22645-206] to Granzyme B -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, IHC-Fr, IPmore details
不适用于: Flow Cyt or ICC/IF -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Mouse splenocyte lysate. IHC-P: Human tonsil and colon tissues; Mouse lung and spleen tissue. IHC-Fr: Mouse spleen tissue; Rat spleen tissue. IP: Mouse spleen tissue.
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常规说明
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR22645-206 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab255598于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
1/1000. Detects a band of approximately 27, 65 kDa (predicted molecular weight: 27 kDa).
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IHC-P | (3) |
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IHC-Fr |
1/200.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
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IP |
1/30.
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说明 |
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WB
1/1000. Detects a band of approximately 27, 65 kDa (predicted molecular weight: 27 kDa). |
IHC-P
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IHC-Fr
1/200. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
IP
1/30. |
靶标
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功能
This enzyme is necessary for target cell lysis in cell-mediated immune responses. It cleaves after Asp. Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -7, -9 and 10 to give rise to active enzymes mediating apoptosis. -
序列相似性
Belongs to the peptidase S1 family. Granzyme subfamily.
Contains 1 peptidase S1 domain. -
细胞定位
Cytoplasmic granule. Cytoplasmic granules of cytolytic T-lymphocytes and natural killer cells. - Information by UniProt
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数据库链接
- Entrez Gene: 3002 Human
- Entrez Gene: 14939 Mouse
- Entrez Gene: 171528 Rat
- Omim: 123910 Human
- SwissProt: P10144 Human
- SwissProt: P04187 Mouse
- SwissProt: P18291 Rat
- Unigene: 1051 Human
see all -
别名
- C11 antibody
- Cathepsin G like 1 antibody
- Cathepsin G-like 1 antibody
see all
图片
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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Granzyme B with ab255598 at 1/3000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes in human tonsil (PMID: 29042640) is observed.
The section was incubated with ab255598 for 30 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Granzyme B with ab255598 at 1/3000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes in human colon is observed.
The section was incubated with ab255598 for 30 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse spleen tissue labeling ab255598 with Granzyme B at 1/200 dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution. The nuclear counterstain was DAPI (Blue).
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
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Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat spleen tissue labeling ab255598 with Granzyme B at 1/200 dilution followed by ab150077 AlexaFluor® 488 Goat anti-Rabbit secondary at 1/1000 dilution. The nuclear counterstain was DAPI (Blue).
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor 488 Goat anti-Rabbit secondary at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Granzyme B with ab255598 at 1/3000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes in mouse lung (PMID: 23642129) is observed.
The section was incubated with ab255598 for 30 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Granzyme B with ab255598 at 1/3000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes in mouse spleen (PMID: 7697916) is observed.
The section was incubated with ab255598 for 30 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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Granzyme B was immunoprecipitated from 0.35 mg mouse spleen cells (treated with 2.5ug/ml Concanavalin A for 72h) whole cell lysate with ab255598 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab255598 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.
Lane 1: Mouse spleen cells (treated with 2.5ug/ml Concanavalin A for 72h) whole cell lysate 10μg
Lane 2: ab255598 IP in mouse spleen cells (treated with 2.5ug/ml Concanavalin A for 72h) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255598 in mouse spleen cells (treated with 2.5ug/ml Concanavalin A for 72h) whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds
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All lanes : Anti-Granzyme B antibody [EPR22645-206] (ab255598) at 1/1000 dilution
Lane 1 : Untreated mouse splenocyte lysate
Lane 2 : Mouse splenocytes treated with 2.5µg/ml concanavalin for 72 hours, lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 27 kDa
Observed band size: 28,65 kDa why is the actual band size different from the predicted?
Exposure time: 8 secondsThe expression of Granzyme B is elevated in stimulated splenocytes. Granzyme B forms complex with its protease inhibitor.
The molecular weight observed is consistent with what has been described in the literature (PMID: 8663264, 17116752, 21919028, 9774654).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (20)
ab255598 被引用在 20 文献中.
- Zhang M et al. Rapamycin improves Graves' orbitopathy by suppressing CD4+ cytotoxic T lymphocytes. JCI Insight 8:N/A (2023). PubMed: 36580373
- Khan SY et al. Tumor-Infiltrating Lymphocytes (TILs) as a Biomarker of Abscopal Effect of Cryoablation in Breast Cancer: A Pilot Study. Ann Surg Oncol 29:2914-2925 (2022). PubMed: 35094188
- Wu SY et al. Combined angiogenesis and PD-1 inhibition for immunomodulatory TNBC: concept exploration and biomarker analysis in the FUTURE-C-Plus trial. Mol Cancer 21:84 (2022). PubMed: 35337339
- Pham TN et al. Inhibition of MNKs promotes macrophage immunosuppressive phenotype to limit CD8+ T cell antitumor immunity. JCI Insight 7:N/A (2022). PubMed: 35380995
- Xu T et al. Cyclin-Dependent Kinase Inhibitors Function as Potential Immune Regulators via Inducing Pyroptosis in Triple Negative Breast Cancer. Front Oncol 12:820696 (2022). PubMed: 35756622