Anti-pan SCN 抗体 [EPR25134-4] (ab300112)

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Immunoprecipitation - Anti-pan SCN antibody [EPR25134-4] (ab300112)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR25134-4] to pan SCN
Suitable for: IHC-P, WB, IHC-Fr, IP
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

製品名

Anti-pan SCN antibody [EPR25134-4]
pan SCN 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR25134-4] to pan SCN
由来種

Rabbit
特異性

This antibody reacts with human SCN1A, SCN3A, and SCN9A but does not react with human SCN4A, SCN5A, SCN8A, SCN10A, and SCN11A.
アプリケーション

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

交差種: Mouse, Rat, Human
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: Human cerebellum, hypothalamus tissue lysates. Mouse brain, hippocampus tissue lysates. Rat brain, hippocampus tissue lysates. HEK-293T cells transfected with a human SCN1A, 2A, 3A, 4A, 5A, 8A , 9A, 10A, and 11A fragment expression vector. IHC-P: Human, mouse, rat brain. IHC-FR: Rat and mouse cerebellum
特記事項
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: 7.20
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

EPR25134-4
アイソタイプ

IgG
研究分野
- Neuroscience
- Processes

Alternative Versions
- Anti-pan SCN antibody [EPR25134-4] (BSA and Azide free) (ab300113)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

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ab300112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
IHC-P		1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/1000. Predicted molecular weight: 228 kDa.
IHC-Fr		1/500.
IP		Use at an assay dependent concentration.

特記事項
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 228 kDa.
IHC-Fr 1/500.
IP Use at an assay dependent concentration.

追加情報

Is unsuitable for Flow Cyt (Intra) or ICC/IF.

Immunoprecipitation - Anti-pan SCN antibody [EPR25134-4] (ab300112)

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Pan SCN was immunoprecipitated from 0.35 mg mouse brain tissue lysate with ab300112 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300112 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse brain tissue lysate 20μg

Lane 2: Mouse brain tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300112 in mouse brain tissue lysate

Exposure time: 10 seconds
Western blot - Anti-pan SCN antibody [EPR25134-4] (AB300112)

All lanes : Anti-pan SCN antibody [EPR25134-4] (ab300112) 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: 228 kDa
Observed band size: 270 kDa why is the actual band size different from the predicted?

Blocking / Diluting buffer and concentration:

5% NFDM/TBST

Exposure time:

5.5 seconds

Negative control: human heart tissue (PMID: 22081212)
Western blot - Anti-pan SCN antibody [EPR25134-4] (AB300112)

All lanes : Anti-pan SCN antibody [EPR25134-4] (ab300112)

Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse hippocampus tissue lysate
Lane 3 : Mouse heart tissue lysate
Lane 4 : Rat brain tissue lysate 20
Lane 5 : Rat hippocampus tissue lysate
Lane 6 : Rat 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: 228 kDa
Observed band size: 270 kDa why is the actual band size different from the predicted?

Exposure time: 26 seconds

Blocking / Diluting buffer and concentration:

5% NFDM/TBST

Negative control: heart (PMID: 22081212)
Western blot - Anti-pan SCN antibody [EPR25134-4] (AB300112)

All lanes : Anti-pan SCN antibody [EPR25134-4] (ab300112) at 1/1000 dilution

Lane 1 : HEK-293T cells transfected with a human SCN1A fragment expression vector containing a his-tag, whole cell lysate
Lane 2 : HEK-293T cells transfected with a human SCN3A fragment expression vector containing a his-tag, whole cell lysate
Lane 3 : HEK-293T cells transfected with a human SCN4A fragment expression vector containing a his-tag, whole cell lysate
Lane 4 : HEK-293T cells transfected with a human SCN5A fragment expression vector containing a his-tag, whole cell lysate
Lane 5 : HEK-293T cells transfected with a human SCN9A fragment expression vector containing a his-tag, 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: 228 kDa
Observed band size: 20-25 kDa why is the actual band size different from the predicted?

Exposure time: 26 seconds

Blocking / Diluting buffer and concentration:

5% NFDM/TBST
Western blot - Anti-pan SCN antibody [EPR25134-4] (AB300112)

All lanes : Anti-pan SCN antibody [EPR25134-4] (ab300112) at 1/1000 dilution

Lane 1 : HEK-293T cells transfected with a human SCN2A expression vector containing a his-tag, whole cell lysate
Lane 2 : HEK-293T cells transfected with a human SCN10A expression vector containing a his-tag, whole cell lysate
Lane 3 : HEK-293T cells transfected with a human SCN11A expression vector containing a his-tag, whole cell lysate
Lane 4 : HEK-293T cells transfected with a human SCN8A expression vector containing a his-tag, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

Predicted band size: 228 kDa
Observed band size: 270 kDa why is the actual band size different from the predicted?

Blocking / Diluting buffer and concentration:

5% NFDM/TBST

Exposure time:

Lane 1-3: 15 seconds
Lane 4: 3 minutes
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Immunohistochemical analysis of paraffin-embedded human brain labeling SCN2A with ab300112 at 1/4000 dilution followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive staining on human brain is observed. The section was incubated with ab300112 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Immunohistochemical analysis of paraffin-embedded mouse brain labeling SCN2A with ab300112 at 1/4000 dilution followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive staining on mouse brain in observed. The section was incubated with ab300112 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling SCN2A with ab300112 at 1/4000 dilution followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive staining on mouse cerebellum is observed. The section was incubated with ab300112 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Immunohistochemical analysis of paraffin-embedded rat brain tissue labeling SCN2A with ab300112 at 1/4000 dilution followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive staining on rat brain is observed. The section was incubated with ab300112 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Frozen sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Positive staining on mouse cerebellum.

Fresh mouse cerebellum was fixed with 4% PFA and permeabilised with 0.2 % Triton X100. Ab300112 was used as a primary antibody at 1/500 dilution. ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was used as a secondary antibody at 1/1000 dilution. DAPI was used as a nuclear counter stain.
Immunohistochemistry (Frozen sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Negative control. No staining on mouse heart

Fresh mouse heart was fixed with 4% PFA and permeabilised with 0.2 % Triton X100. Ab300112 was used as a primary antibody at 1/500 dilution. ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was used as a secondary antibody at 1/1000 dilution. DAPI was used as a nuclear counter stain.
Immunohistochemistry (Frozen sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Positive staining on Rat cerebellum.

Fresh Rat cerebellum was fixed with 4% PFA and permeabilised with 0.2 % Triton X100. Ab300112 was used as a primary antibody at 1/500 dilution (0.862 μg/ml). ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was preadsorbed and used as a secondary antibody at 1/1000 dilution (2 μg/ml). DAPI was used as a nuclear counter stain.
Immunohistochemistry (Frozen sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Negative control. No staining on rat heart

Fresh rat heart was fixed with 4% PFA and permeabilised with 0.2 % Triton X100. Ab300112 was used as a primary antibody at 1/500 dilution. ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was used as a secondary antibody at 1/1000 dilution. DAPI was used as a nuclear counter stain.
Anti-pan SCN antibody [EPR25134-4] (AB300112)

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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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

特異性

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

アプリケーション

The Abpromise guarantee

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 (0)

レビューと Q&A