Anti-SPHK1 抗体 [EPR24511-37] (ab302714)

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Western blot - Anti-SPHK1 antibody [EPR24511-37] (AB302714)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR24511-37] to SPHK1
Suitable for: IP, WB
Reacts with: Human

Carrier Free

製品名

Anti-SPHK1 antibody [EPR24511-37]
SPHK1 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR24511-37] to SPHK1
由来種

Rabbit
アプリケーション

適用あり: IP, WBmore details
適用なし: Flow Cyt (Intra),ICC/IF or IHC-P
種交差性

交差種: Human
非交差種: Mouse, Rat
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: Whole cell lysates: Hela (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control, Hela transfected with siRNA targeting SPHK1, U-118 MG (human brain glioma cell), SW1353 (human chondrosarcoma fibroblast), HeLa, HepG2 (human hepatocellular carcinoma epithelial cell). Tissue lysates: Human kidney, heart, lung. IP: Whole cell lysates: U-118 MG (human brain glioblastoma cell), HepG2 (human hepatocellular carcinoma epithelial cell).
特記事項
ab302714 does not react in: WB and immunoprecipitation with mouse and rat species: IHC-P, ICC, and intracellular flow cytometry with human, mouse, and rat species.

This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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 long term. Avoid freeze / thaw cycle.
バッファー

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR24511-37
アイソタイプ

IgG
研究分野

Alternative Versions
- Anti-SPHK1 antibody [EPR24511-37] (BSA and Azide free) (ab302715)
Compatible Secondaries
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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ab302714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
IP		1/30.
WB		1/1000. Detects a band of approximately 42-52 kDa (predicted molecular weight: 43 kDa).

特記事項
IP 1/30.
WB 1/1000. Detects a band of approximately 42-52 kDa (predicted molecular weight: 43 kDa).

追加情報

Is unsuitable for Flow Cyt (Intra),ICC/IF or IHC-P.

機能

Catalyzes the phosphorylation of sphingosine to form sphingosine 1-phosphate (SPP), a lipid mediator with both intra-and extracellular functions. Also acts on D-erythro-sphingosine and to a lesser extent sphinganine, but not other lipids, such as D,L-threo-dihydrosphingosine, N,N-dimethylsphingosine, diacylglycerol, ceramide, or phosphatidylinositol.
組織特異性

Widely expressed with highest levels in adult liver, kidney, heart and skeletal muscle.
配列類似性

Contains 1 DAGKc domain.
Target information above from: UniProt accession Q9NYA1 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 8877 Human
- Omim: 603730 Human
- SwissProt: Q9NYA1 Human
- Unigene: 68061 Human
別名
- SK 1 antibody
- SK1 antibody
- Sphingosine kinase 1 antibody
- SPHK 1 antibody
- SPHK antibody
- Sphk1 antibody
- SPHK1_HUMAN antibody
- SPK antibody
- SPK 1 antibody
see all

Western blot - Anti-SPHK1 antibody [EPR24511-37] (AB302714)

All lanes : Anti-SPHK1 antibody [EPR24511-37] (ab302714) at 1/1000 dilution

Lane 1 : Hela (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control, whole cell lysate
Lane 2 : Hela transfected with siRNA specifically targeting SPHK1, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 43 kDa
Observed band size: 42-52 kDa why is the actual band size different from the predicted?

Exposure time: 180 seconds

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

he additional band at ~50 kDa is expected to be isoform 2 (Q9NYA1-2) of SPHK1 and its expression is also significantly reduced through SPHK1 knockdown.
Western blot - Anti-SPHK1 antibody [EPR24511-37] (AB302714)

All lanes : Anti-SPHK1 antibody [EPR24511-37] (ab302714) at 1/1000 dilution

Lane 1 : Human kidney tissue lysate
Lane 2 : Human heart tissue lysate
Lane 3 : Human lung 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 1/1000 dilution

Predicted band size: 43 kDa
Observed band size: 4-52 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: Lane 1 and 2: 180 seconds, Lane 3: 15 seconds
Western blot - Anti-SPHK1 antibody [EPR24511-37] (AB302714)

All lanes : Anti-SPHK1 antibody [EPR24511-37] (ab302714) at 1/1000 dilution

Lane 1 : U-118 MG (human brain glioma cell), whole cell lysate
Lane 2 : SW1353 (human chondrosarcoma fibroblast), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 43 kDa
Observed band size: 42-52 kDa why is the actual band size different from the predicted?

Exposure time: 70 seconds

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Western blot - Anti-SPHK1 antibody [EPR24511-37] (AB302714)

All lanes : Anti-SPHK1 antibody [EPR24511-37] (ab302714) at 1/1000 dilution

Lane 1 : Hela (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 43 kDa
Observed band size: 42-52 kDa why is the actual band size different from the predicted?

Exposure time: 70 seconds

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Immunoprecipitation - Anti-SPHK1 antibody [EPR24511-37] (AB302714)

SPHK1 was immunoprecipitated from 0.35 mg U-118 MG (human brain glioblastoma cell) whole cell lysate with ab302714 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302714 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: U-118 MG (human brain glioblastoma cell) whole cell lysate 10ug

Lane 2: ab302714 IP in U-118 MG whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302714 in U-118 MG whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 180 seconds

Observed MW (kDa): 42-52
Immunoprecipitation - Anti-SPHK1 antibody [EPR24511-37] (AB302714)

SPHK1 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab302714 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302714 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug

Lane 2: ab302714 IP in HepG2 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302714 in HepG2 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 180 seconds

Observed MW (kDa): 42-52
Anti-SPHK1 antibody [EPR24511-37] (AB302714)

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.

Click here to view the general protocols

SDS download

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Datasheet download

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ab302714 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab302714 は 1 報の論文で使用されています。

Xu J et al. Transcriptome and Lipidomic Analysis Suggests Lipid Metabolism Reprogramming and Upregulating SPHK1 Promotes Stemness in Pancreatic Ductal Adenocarcinoma Stem-like Cells. Metabolites 13:N/A (2023). PubMed: 37999228

レビューと Q&A

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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

配列類似性

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

Certificate of Compliance

参考文献 (1)

レビューと Q&A