Anti-Caspr 抗体 (ab34151)
Key features and details
- Rabbit polyclonal to Caspr
- Suitable for: WB, ICC/IF, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-Caspr antibody
Caspr 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Caspr -
由来種
Rabbit -
アプリケーション
適用あり: WB, ICC/IF, IPmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide corresponding to Mouse Caspr aa 1350 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab34150) -
ポジティブ・コントロール
- ICC/IF: Primary rat neurons/glia, DIV14 cells. SKNSH cells. WB: Wild type HAP1 whole cell lysate. MEF and HeLa whole cell lysate. Rat brain whole cell lysate. IP: Rat brain whole cell lysate.
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
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. -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab34151の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
1/250. Detects a band of approximately 180 kDa (predicted molecular weight: 156 kDa).Can be blocked with Mouse Caspr peptide (ab34150).
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ICC/IF |
Use a concentration of 1 µg/ml.
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IP |
Use at an assay dependent concentration.
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特記事項 |
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WB
1/250. Detects a band of approximately 180 kDa (predicted molecular weight: 156 kDa).Can be blocked with Mouse Caspr peptide (ab34150). |
ICC/IF
Use a concentration of 1 µg/ml. |
IP
Use at an assay dependent concentration. |
ターゲット情報
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機能
Seems to play a role in the formation of functional distinct domains critical for saltatory conduction of nerve impulses in myelinated nerve fibers. Seems to demarcate the paranodal region of the axo-glial junction. In association with contactin may have a role in the signaling between axons and myelinating glial cells. -
組織特異性
Predominantly expressed in brain. Weak expression detected in ovary, pancreas, colon, lung, heart, intestine and testis. -
配列類似性
Belongs to the neurexin family.
Contains 2 EGF-like domains.
Contains 1 F5/8 type C domain.
Contains 1 fibrinogen C-terminal domain.
Contains 4 laminin G-like domains. -
細胞内局在
Membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 8506 Human
- Entrez Gene: 53321 Mouse
- Entrez Gene: 84008 Rat
- Omim: 602346 Human
- SwissProt: P78357 Human
- SwissProt: O54991 Mouse
- SwissProt: P97846 Rat
- Unigene: 408730 Human
see all -
別名
- Caspr antibody
- Caspr1 antibody
- CNTNAP antibody
see all
画像
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ab34151 staining Caspr in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween 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 overnight at 4°C with ab34151 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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ab34151 stained in SKNSH cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab34151 at 1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
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All lanes : Anti-Caspr antibody (ab34151) at 1/250 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CNTNAP1 (Caspr) knockout HAP1 whole cell lysate
Lane 3 : MEF whole cell lysate
Lane 4 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 156 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab34151 observed at 180 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab34151 was shown to recognize Caspr in wild-type HAP1 cells as signal was lost at the expected MW in CNTNAP1 (Caspr) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CNTNAP1 (Caspr) knockout samples were subjected to SDS-PAGE. ab34151 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/250 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Anti-Caspr antibody (ab34151) at 1/250 dilution + Brain (Rat) Whole Cell Lysate - normal tissue at 10 µg
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 156 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?
Additional bands at: 58 kDa. We are unsure as to the identity of these extra bands.
Caspr contains a number of potential glycosylation sites so it is thought that this is the reason it runs at 180kDa. -
Caspr was immunoprecipitated using 0.5mg Rat Brain whole tissue lysate, 5µg of Rabbit polyclonal to Caspr and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Rat Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab34151.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 180kDa: Caspr.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (72)
ab34151 は 72 報の論文で使用されています。
- Ishino Y et al. Coactivator-associated arginine methyltransferase 1 controls oligodendrocyte differentiation in the corpus callosum during early brain development. Dev Neurobiol 82:245-260 (2022). PubMed: 35220691
- Joost S et al. Cuprizone Intoxication Results in Myelin Vacuole Formation. Front Cell Neurosci 16:709596 (2022). PubMed: 35250482
- Jordans S et al. Age-Dependent Increase in Schmidt-Lanterman Incisures and a Cadm4-Associated Membrane Skeletal Complex in Fatty Acid 2-hydroxylase Deficient Mice: a Mouse Model of Spastic Paraplegia SPG35. Mol Neurobiol 59:3969-3979 (2022). PubMed: 35445918
- Radomski KL et al. Acute axon damage and demyelination are mitigated by 4-aminopyridine (4-AP) therapy after experimental traumatic brain injury. Acta Neuropathol Commun 10:67 (2022). PubMed: 35501931
- Mo YJ et al. Advantages of Adult Mouse Dorsal Root Ganglia Explant Culture in Investigating Myelination in an Inherited Neuropathic Mice Model. Methods Protoc 5:N/A (2022). PubMed: 35893592