Anti-Sortilin/NT3抗体
参阅全部 Sortilin/NT3 一抗
兔多克隆抗体to Sortilin/NT3
Rabbit
适用于: IHC-FoFr, WBmore details
与反应: Mouse, Rat, Human
预测可用于: Cow
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
WB: HeLa cell lysate; NIH/3T3 cell lysate; 293T cell lysate; Human, Mouse, and Rat Brain tissue lysates. IHC-FoFr: Rat cortical neurons.
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
浓度
批次浓度范围 100 µg 浓度为 0.7 - 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab16640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 |
|---|---|---|
| IHC-FoFr | (2) | 1/3000. |
| WB | (5) | Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa). |
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Unigene: 703487 Human
Unigene: 157119 Mouse
Unigene: 11286 Rat
Entrez Gene: 6272 Human
Entrez Gene: 20661 Mouse
Omim: 602458 Human
SwissProt: Q99523 Human
SwissProt: Q6PHU5 Mouse
Unigene: 485195 Human
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Western blot - Anti-Sortilin/NT3 antibody (ab16640)
All lanes : Anti-Sortilin/NT3 antibody (ab16640) at 1 µg/ml
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SORT1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 95 kDa
Observed band size: 100 kDawhy is the actual band size different from the predicted?
Lanes 1- 2: Merged signal (red and green). Green - ab16640 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab16640 was shown to react with Sortilin/NT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264772 (knockout cell lysate ab257696) was used. Wild-type HeLa and SORT1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab16640 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Sortilin/NT3 antibody (ab16640)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Sortilin/NT3 knockout HAP1 cell lysate (20 µg)
Lane 3: NIH/3T3 cell lysate (20 µg)
Lane 4: 293T cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab16440 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab16640 was shown to specifically react with Sortilin/NT3 in wild-type HAP1 cells. No band was observed when Sortilin/NT3 knockout samples were examined. Wild-type and Sortilin/NT3 knockout samples were subjected to SDS-PAGE. ab16640 and ab8245 (loading control to GAPDH) were diluted 1 µg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Sortilin/NT3 antibody (ab16640)
Lane 1 : Marker
Lanes 2-4 : Anti-Sortilin/NT3 antibody (ab16640) at 1 µg/ml
Lane 2 : Human Brain tissue lysate at 20 µg
Lane 3 : Mouse Brain tissue lysate at 20 µg
Lane 4 : Rat Brain whole cell lysate at 20 µg
Secondary
Lanes 2-4 : Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 95 kDa
Additional bands at: 31 kDa (possible cleavage fragment)
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Sortilin/NT3 antibody (ab16640)This image is courtesy of Sophie Pezet, Univ London Kings Coll, United Kingdom
Immunofluorescent staining for Sortilin/NT3 in rat cortical neurons using ab16640. The staining is cytoplasmic and punctate of many cells such as cortical neurons. The image was taken with a X20 objective.
Protocol details: Rats were intracardially perfused with 4% paraformaldehyde. Whole brain tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT and then cut on cryostat (30µm coronal sections). IHC was perfored in free floating with fixed tissues (rat brain sections). Primary antibody was incubated overnight at 1/3000 at room temperature
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