TNFSF9 抗体 (AA 101-200)
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- 抗原 See all TNFSF9 抗体
- TNFSF9 (Tumor Necrosis Factor (Ligand) Superfamily, Member 9 (TNFSF9))
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抗原表位
- AA 101-200
- 适用
- 人, 小鼠, 大鼠
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宿主
- 兔
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克隆类型
- 多克隆
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标记
- This TNFSF9 antibody is un-conjugated
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应用范围
- Western Blotting (WB), ELISA, Immunocytochemistry (ICC), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))
- 交叉反应
- 人, 小鼠, 大鼠
- 纯化方法
- Purified by Protein A.
- 免疫原
- KLH conjugated synthetic peptide derived from human TNFSF9
- 亚型
- IgG
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- 应用备注
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WB 1:300-5000
ELISA 1:500-1000
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200
ICC 1:100-500 - 限制
- 仅限研究用
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- by
- Immunohistochemistry Core, NYU Langone
- No.
- #029578
- 日期
- 2014.01.18
- 抗原
- Lot Number
- 999892W
- Method validated
- Immunohistochemistry
- Positive Control
- Human liver
- Negative Control
- Human placental decidual cells
- Notes
- Signal was detected in positive control tissue and not in negative control tissue.
- Primary Antibody
- Antibody: Tumor necrosis factor ligand superfamily member 9 (TNFSF9)
- Catalog number: ABIN705056
- Lot number: 999892W
- Secondary Antibody
- Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
- Lot number: D05923BA
- Full Protocol
- Immunohistochemistry was performed on a Ventana NexES automated platform, instrument manufacturer specific reagents are italicized.
- 1. Slides were preheated in convection oven at 60°C for 30 minutes
- 2. Deparaffinization procedure: - 3 changes of Xylene, 5 minutes each - 3 changes of 100% Ethanol, 3 minutes each - 3 changes of 95% Ethanol, 3 minutes each - Rinsed in distilled water, 3 changes
- 1. Heat retrieval procedure - Slides retrieved in 10.0 mM Citrate, pH6.0 in a 1000W microwave oven (~100°C) for 15 minutes. - Slides were allowed to cool (in citrate) for 30 minutes. - Slides were washed x 3 in Distilled water
- 1. NexES instrument procedure, iVIEW DAB paraffin protocol (*abridged*): - Slide chamber warmed to 37°C
- 1. Slides rinsed with *reaction buffer* x 3
- 1. *iVIEW Inhibitor (H2O2)* applied and incubated for 4 minutes
- 1. Slides rinsed with *reaction buffer*
- 1. Antibody Application - Primary antibody diluted 1:250 in PBS (100 microliters applied/slide) - Ventana Isotype control applied neat - Slides incubated overnight at room temperature (~12 hours ~25°C)
- 1. Slides rinsed with *reaction buffer* x3
- 1. *iVIEW Biotinylated IgG* applied and incubated for 8 minutes
- 1. Slides rinsed with *reaction buffer*
- 1. *iVIEW Streptavidin-Horseradish Peroxidase* applied and incubated for 8 minutes
- 1. Slides rinsed with *reaction buffer*
- 1. *iVIEW DAB/H2O2* applied and incubated for 8 minutes
- 1. Slides rinsed with *reaction buffer*
- 1. *iVIEW Copper* applied and incubated for 4 minutes
- 1. Slides rinsed with *reaction buffer*
- 1. Slides washed in Dawn Detergent/tap water
- 1. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 seconds - Slides washed in tap water, 1 minute - Decolorized (10% Acetic Acid in 70% ethanol), 1 minute - Slides washed in tap water, 1 minute - Bluing (Austin Clear Ammonia), 1 minute - Slides washed in tap water, 1 minute
- 1. Dehydration/coverslipping procedure: - 3 changes of 95% Ethanol, 3 minutes each - 3 changes of 100% Ethanol, 3 minutes each - 3 changes of Xylene, 5 minutes each - Mounted with Permount
- 1. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software
- Experimental Notes
- Deviations from protocol/procedure supplied by manufacturer:
- Step 1: Heated tissue 60°C for 30 minutes; manufacturer heats for 45 minutes.
- Step 2: No ethanol wash was performed during deparaffinization; manufacturer includes 1 wash of 80% ethanol for 3 minutes.
- Step 3.1: Slides heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
- Step 3.2: Slides cooled for 30 minutes; manufacturer cools for 20 minutes.
- Step 4: Italicized reagents and incubation time are fixed instrument parameters.
- Step 5: Secondary species-specific serum block not used; manufacturer blocks with 5% normal goat serum for 2 hours.
- Step 8.1: Antibody diluted in PBS at 1:250; manufacture recommends dilution range (1:100-1:500). No primary antibody diluent recommended.
- Step 8.2.1: Primary antibody incubated at room temperature overnight; manufacturer incubates overnight 4°C with agitation.
- Tissue Interpretation (limited): • TNFSF9: Under the staining parameters described above, placenta was weakly cytoplasmic positive (trophoblastic and decidua cells). Liver hepatocytes were weakly stained. Substantial signal detected in limited number of other tissues, including kidney, prostate, and skin.
- • I-NC (Isotype negative control): No signal detected
- • B-NC (Blank negative control): No signal detected
- Signal Localization: - Signal to noise was adequate with predominately cytoplasmic subcellular localization observed. Nuclear staining was rare and no distinct membrane signal was detected.
生效 #029578 (Immunohistochemistry)Validation ImagesFull Methods -
- 状态
- Liquid
- 浓度
- 1 μg/μL
- 缓冲液
- 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.
- 储存液
- ProClin
- 注意事项
- This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
- 储存条件
- 4 °C,-20 °C
- 储存方法
- Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
- 有效期
- 12 months
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- 抗原
- TNFSF9 (Tumor Necrosis Factor (Ligand) Superfamily, Member 9 (TNFSF9))
- 别名
- TNFSF9/CD137L (TNFSF9 产品)
- 别名
- TNFSF9 antibody, 4-1BB-L antibody, 4-1BBL antibody, AI848817 antibody, Cd137l antibody, Ly63l antibody, CD137L antibody, TNF superfamily member 9 antibody, tumor necrosis factor (ligand) superfamily, member 9 antibody, Tnfsf9 antibody, LOC476729 antibody, TNFSF9 antibody
- 背景
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Synonyms: CD137L, 4-1BB-L, Tumor necrosis factor ligand superfamily member 9, 4-1BB ligand, 4-1BBL, TNFSF9
Background: Cytokine that binds to TNFRSF9. Induces the proliferation of activated peripheral blood T-cells. May have a role in activation-induced cell death (AICD). May play a role in cognate interactions between T-cells and B-cells/macrophages.
- 基因ID
- 8744
- UniProt
- P41273
- 途径
- Activated T Cell Proliferation, Cancer Immune Checkpoints
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