重组Anti-VCAM1抗体[EPR5047] (ab134047)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5047] to VCAM1
- Suitable for: WB, IP, IHC-P, Flow Cyt (Intra), ICC/IF, Indirect ELISA
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-VCAM1抗体[EPR5047]
参阅全部 VCAM1 一抗 -
描述
兔单克隆抗体[EPR5047] to VCAM1 -
宿主
Rabbit -
经测试应用
适用于: WB, IP, IHC-P, Flow Cyt (Intra), ICC/IF, Indirect ELISAmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
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阳性对照
- IHC-P: Human spleen and tonsil tissue; mouse spleen tissue. WB: Mouse kidney, brain and spleen tissue lysate; rat brain, spleen and kidney tissue lysate; human fetal liver tissue lysate; NIH/3T3, LADMAC, HuT-78, TNF-a treated HUVEC, and LPS treated bEnd.3 cell lysates; Wild-type A549 and HUVEC TNF-a treated (10 ng/mL, 16h) cell lysate Flow Cyt (intra): K562 cells. ICC/IF: HUVEC TNF-a treated (10 ng/mL, 16h); K562 cells. IP: Human fetal liver tissue lysate.
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常规说明
Vascular cell adhesion protein 1 (VCAM) is a protein that is encoded by the VCAM1 gene in humans. It plays a role in functioning as a cell adhesion molecule and is thought to participate in monocyte recruitment to atherosclerotic sites, and as such is highly overexpressed in brain inflammation.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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR5047 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 647 Anti-VCAM1 antibody [EPR5047] (ab194319)
- HRP Anti-VCAM1 antibody [EPR5047] (ab195540)
- Anti-VCAM1 antibody [EPR5047] - Low endotoxin, Azide free (ab215380)
- PE Anti-VCAM1 antibody [EPR5047] (ab223981)
- FITC Anti-VCAM1 antibody [EPR5047] (ab223982)
- APC Anti-VCAM1 antibody [EPR5047] (ab223983)
- Anti-VCAM1 antibody [EPR5047] - BSA and Azide free (ab271899)
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab134047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 | (3) |
1/2000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 81 kDa).
Stimulation may be required to allow detection of the target protein due to low levels of endogenous expression in some samples. Please see images below for recommended treatment conditions and positive controls. |
IP |
1/40.
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IHC-P | (5) |
1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/40.
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ICC/IF | (3) |
1/250.
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Indirect ELISA |
Use at an assay dependent concentration.
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说明 |
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WB
1/2000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 81 kDa). Stimulation may be required to allow detection of the target protein due to low levels of endogenous expression in some samples. Please see images below for recommended treatment conditions and positive controls. |
IP
1/40. |
IHC-P
1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/40. |
ICC/IF
1/250. |
Indirect ELISA
Use at an assay dependent concentration. |
靶标
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功能
Important in cell-cell recognition. Appears to function in leukocyte-endothelial cell adhesion. Interacts with the beta-1 integrin VLA4 on leukocytes, and mediates both adhesion and signal transduction. The VCAM1/VLA4 interaction may play a pathophysiologic role both in immune responses and in leukocyte emigration to sites of inflammation. -
组织特异性
Expressed on inflammed vascular endothelium, as well as on macrophage-like and dendritic cell types in both normal and inflammed tissue. -
序列相似性
Contains 7 Ig-like C2-type (immunoglobulin-like) domains. -
结构域
Either the first or the fourth Ig-like C2-type domain is required for VLA4-dependent cell adhesion. -
翻译后修饰
Sialoglycoprotein. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 7412 Human
- Entrez Gene: 22329 Mouse
- Entrez Gene: 25361 Rat
- GenBank: NP_001069.1 Human
- GenBank: NP_001186763.1 Human
- Omim: 192225 Human
- SwissProt: P19320 Human
- SwissProt: P29533 Mouse
see all -
别名
- CD106 antibody
- CD106 Antigen antibody
- INCAM 100 antibody
see all
图片
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ELISA using ab134047 at varying antibody concentrations and antigen concentration at 250 ng/mL. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
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All lanes : Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/2000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate
Lane 3 : VCAM1 knockout A549 cell lysate
Lane 4 : VCAM1 knockout A549 TNF-a treated (10 ng/mL, 16h) cell lysate
Lane 5 : HUVEC cell lysate
Lane 6 : HUVEC TNF-a treated (16 ng/mL, 16h) cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?Lanes 1 - 6: Merged signal (red and green). Green - ab134047 observed at 105 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab134047 was shown to react with VCAM1 in treated wild-type A549 cells in Western blot with loss of signal observed in treated VCAM1 knockout cell line ab273758 (knockout cell lysate ab275504). Wild-type A549 and VCAM1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab134047 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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ab134047 staining VCAM1 in HUVEC cells treated with TNF-α (ab55237) at 10 ng/ml for 16 hours. The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab134047 at 2μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 647) (ab150119) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
All lanes : Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/1000 dilution
Lane 1 : HUVEC (Human umbilical vein endothelial cell) whole cell lysate
Lane 2 : HUVEC (Human umbilical vein endothelial cell) treated with 10 ng/ml TNF-a for 16 h, whole cell lysate
Lane 3 : bEnd.3 (Mouse brain endothelioma) whole cell lysate
Lane 4 : bEnd.3 (Mouse brain endothelioma) treated with 10 µg/ml LPS for 24 h, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Additional bands at: 36 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 secondsRabbit monoclonal [EPR16891] to GAPDH (ab181602) used as loading control.
Blocking/Diluting buffer: 5% NFDM/TBST.
Stimulation may be required to allow detection of the target protein due to low levels of endogenous expression in some samples.
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All lanes : Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/1000 dilution
Lane 1 : Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate
Lane 2 : SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysate
Lane 3 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 4 : J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate
Lane 5 : LADMAC (Mouse bone marrow monocyte macrophage) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Additional bands at: 36 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 7 secondsRabbit monoclonal [EPR16891] to GAPDH (ab181602) used as loading control.
Blocking/Diluting buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCAM1 antibody [EPR5047] (ab134047)
Immunohistochemical staining of paraffin embedded human tonsil with purified ab134047 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunofluorescent staining of K562 cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab134047 at a dilution of 1/250. An Alexa Fluor® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counter stained with DAPI. The negative control is shown in the bottom right hand panel - for the negative control, Alex Fluor® 594 goat anti-mouse was used at a dilution of 1/500.
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Intracellular Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) cells labeling VCAM1 with purified ab134047 at 1/40 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
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All lanes : Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/10000 dilution (purified)
Lane 1 : Mouse kidney
Lane 2 : Rat spleen
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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VCAM1 was immunoprecipitated from Human fetal liver lysate with ab134047 at 1/110 dilution.
Western blot was performed from the immunoprecipitate using ab134047 at 1/1000 dilution.
VeriBlot for IP secondary antibody (Peroxidase conjugated),was used as secondary antibody at 1/1000 dilution.
Lane 1: Human fetal liver lysate.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 1 second.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCAM1 antibody [EPR5047] (ab134047)
IHC image of VCAM1 staining in Human spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with unpurified ab134047, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin 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.
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Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/2000 dilution (purified) + Rat kidney tissue lysate at 20 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM/TBST.
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Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/10000 dilution (purified) + NIH/3T3 cell lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM/TBST.
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Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/10000 dilution (purified) + Human fetal liver tissue lysate at 20 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM/TBST.
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Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/10000 dilution (purified) + HuT-78 cell lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCAM1 antibody [EPR5047] (ab134047)
IHC image of VCAM1 staining in Mouse spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with unpurified ab134047, 1/200 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin 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.
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All lanes : Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/1000 dilution (unpurified)
Lane 1 : Human fetal liver tissue lysate
Lane 2 : HuT 78 cell lysate
Lane 3 : NIH/3T3 cell lysate
Lane 4 : Mouse brain tissue lysate
Lane 5 : Mouse kidney tissue lysate
Lane 6 : Mouse spleen tissue lysate
Lane 7 : Rat brain tissue lysate
Lane 8 : Rat kidney tissue lysate
Lane 9 : Rat spleen tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution
Predicted band size: 81 kDa
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (258)
ab134047 被引用在 258 文献中.
- Qiao T et al. Combined pembrolizumab and bevacizumab therapy effectively inhibits non-small-cell lung cancer growth and prevents postoperative recurrence and metastasis in humanized mouse model. Cancer Immunol Immunother 72:1169-1181 (2023). PubMed: 36357599
- Baganha F et al. Phosphorylcholine Monoclonal Antibody Therapy Decreases Intraplaque Angiogenesis and Intraplaque Hemorrhage in Murine Vein Grafts. Int J Mol Sci 23:N/A (2022). PubMed: 36362449
- Wu J et al. Vessel state and immune infiltration of the angiogenesis subgroup and construction of a prediction model in osteosarcoma. Front Immunol 13:992266 (2022). PubMed: 36405691
- Ackerman JE et al. Defining the spatial-molecular map of fibrotic tendon healing and the drivers of Scleraxis-lineage cell fate and function. Cell Rep 41:111706 (2022). PubMed: 36417854
- Mastrogiacomo L & Werstuck GH Investigating the Role of Endothelial Glycogen Synthase Kinase3α/β in Atherogenesis in Low Density Lipoprotein Receptor Knockout Mice. Int J Mol Sci 23:N/A (2022). PubMed: 36499109