重组Anti-RILPL1抗体[MJF-R41-21] (ab302492)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [MJF-R41-21] to RILPL1
- Suitable for: ICC/IF, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-RILPL1抗体[MJF-R41-21]
参阅全部 RILPL1 一抗 -
描述
兔单克隆抗体[MJF-R41-21] to RILPL1 -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, IP, WBmore details
不适用于: Flow Cyt,IHC-Fr or IHC-P -
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: Wild-type A549 whole cell lysate. HeLa, NIH/3T3, C6, SH-SY5Y, HEK-293, Neuro-2a, and PC-12 whole cell lysates; rat and mouse brain tissue lysate. Human cerebellum, hypothalamus and heart tissue lysate. ICC: SH-SY5Y and Neuro-2a cells. IP: A549 whole cell lysate, mouse and rat brain tissue lysates.
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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.
This antibody was developed with support from The Michael J. Fox Foundation.
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
MJF-R41-21 -
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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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab302492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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ICC/IF |
1/250.
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IP |
1/30.
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WB |
1/1000. Detects a band of approximately 47, 42 kDa (predicted molecular weight: 47 kDa).
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说明 |
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ICC/IF
1/250. |
IP
1/30. |
WB
1/1000. Detects a band of approximately 47, 42 kDa (predicted molecular weight: 47 kDa). |
靶标
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功能
Neuroprotective protein, which acts by sequestring GAPDH in the cytosol and prevent the apoptotic function of GAPDH in the nucleus. Competes with SIAH1 for binding GAPDH (By similarity). Does not regulate lysosomal morphology and distribution. -
组织特异性
Widely expressed. Expressed at lower level in liver and kidney. -
序列相似性
Belongs to the RILPL family.
Contains 1 RILP-like domain. -
翻译后修饰
S-nitrosylation is required for the interaction with GAPDH. -
细胞定位
Cytoplasm > cytosol. - Information by UniProt
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数据库链接
- Entrez Gene: 353116 Human
- Entrez Gene: 75695 Mouse
- Entrez Gene: 304469 Rat
- GenBank: NM_178314.3 Human
- Omim: 614092 Human
- SwissProt: Q5EBL4 Human
- SwissProt: Q9JJC6 Mouse
- SwissProt: D3ZUQ0 Rat
see all -
别名
- GOSPEL antibody
- MGC105128 antibody
- MGC99793 antibody
see all
图片
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All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution
Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell), whole cell lysate
Lane 2 : RILPL1 knockout A549 whole cell lysate
Lane 3 : HeLa (human cervical adenocarcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 47 kDa
Observed band size: 42,47 kDa why is the actual band size different from the predicted?Blocking / Diluting buffer and: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
False colour image of Western blot: Anti-RILPL1 antibody [MJF-R41-21] (ab302492) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab302492 was shown to bind specifically to RILPL1. Two bands were observed at 47/42 kDa in wild-type A549 cell lysates whereas no signal observed at this size in RILPL1 knockout cell line. To generate this image, wild-type and RILPL1 knockout A549 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel and then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.
The doublet bands represent two isoforms of RILPL1.
Wild-type and RILPL1 knockout A549 cell lysates were kindly provided by Dr. Dario Alessi, University of Dundee. -
All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution
Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell), whole cell lysate
Lane 2 : RILPL1 knockout A549 whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 5 : Rat brain tissue lysate
Lane 6 : Rat kidney tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 47 kDa
Observed band size: 42,47 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking / Diluting buffer and: 5% NFDM/TBST.
The doublet bands represent two isoforms of RILPL1.
Negative control: kidney (PMID: 14668488).
Wild-type and RILPL1 knockout A549 cell lysates were kindly provided by Dr. Dario Alessi, University of Dundee. -
All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lane 3 : Mouse liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: 103 secondsBlocking / Diluting buffer and: 5% NFDM/TBST.
Negative control: kidney, liver (PMID: 14668488).
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All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution
Lane 1 : SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate
Lane 2 : U937 (human histiocytic lymphoma monocyte), whole cell lysate
Lane 3 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 47 kDa
Observed band size: 42,47 kDa why is the actual band size different from the predicted?
Exposure time: 37 secondsBlocking / Diluting buffer and: 5% NFDM/TBST.
The doublet bands represent two isoforms of RILPL1.
Low expression: U937 (Human Protein Atlas). -
All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution
Lane 1 : Human cerebellum tissue lysate
Lane 2 : Human hypothalamus tissue lysate
Lane 3 : Human heart tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 47 kDa
Observed band size: 42,47 kDa why is the actual band size different from the predicted?
Exposure time: 26 secondsBlocking / Diluting buffer and: 5% NFDM/TBST.
The doublet bands represent two isoforms of RILPL1.
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Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) labelling RILPL1 with ab302492 at 1/250 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (green). Confocal image showing centrosome and cytoplasmic staining in SH-SY5Y cell line. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (red) at 1/200 dilution.
Low expression: U-937
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Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) labelling RILPL1 with ab302492 at 1/250 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (green). Confocal image showing centrosome staining in Neuro-2a cell line. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (red) at 1/200 dilution.
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RILPL1 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell), whole cell lysate with ab302492 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302492 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1 (Input): A549 whole cell lysate 10 μg
Lane 2 (+): A549 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab302492 in A549 whole cell lysate
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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RILPL1 was immunoprecipitated from 0.35 mg mouse brain tissue lysate with ab302492 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302492 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1 (Input): Mouse brain tissue lysate 10 μg
Lane 2 (+): Mouse brain tissue lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab302492 in mouse brain tissue lysate
Blocking and dilution buffer and: 5% NFDM/TBST.
Exposure time: 3 minutes.
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RILPL1 was immunoprecipitated from 0.35 mg rat brain tissue lysate with ab302492 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302492 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1 (Input): Rat brain tissue lysate 10 μg
Lane 2 (+): Rat brain tissue lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab302492 in rat brain tissue lysate
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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 (0)
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