Anti-ADNP 抗体 [EPR25434-4] - BSA and Azide free (ab300115)

製品名

Anti-ADNP antibody [EPR25434-4] - BSA and Azide free
ADNP 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR25434-4] to ADNP - BSA and Azide free
由来種

Rabbit
アプリケーション

適用あり: WB, IHC-P, IHC-Fr, Flow Cyt (Intra), IP, ICC/IFmore details
適用なし: ChIP
種交差性

交差種: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: 293T, NIH/3T3, PC-12, HeLa, C6, U-87 MG, SH-SY5Y, Neuro-2a, mouse brain and rat brain lysates. IHC-P: Human colon carcinoma, mouse cerebrum, mouse glioblastoma and rat cerebrum tissues. IHC-Fr: Mouse cerebrum and rat cerebrum tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt. Intra.: HeLa and NIH/3T3 cells. IP: HeLa and NIH/3T3 cells.
特記事項
Ab300115 is a carrier free version of ab300114.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.
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.
バッファー

pH: 7.2
Constituent: 100% PBS
キャリア・フリー

はい
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

EPR25434-4
アイソタイプ

IgG
研究分野
- Neuroscience
- Development

Alternative Versions
- Anti-ADNP antibody [EPR25434-4] (ab300114)
Compatible Secondaries
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ab300115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
WB		Use at an assay dependent concentration. Predicted molecular weight: 124 kDa.
IHC-P		Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr		Use at an assay dependent concentration.
Flow Cyt (Intra)		Use at an assay dependent concentration.
IP		Use at an assay dependent concentration.
ICC/IF		Use at an assay dependent concentration.

特記事項
WB Use at an assay dependent concentration. Predicted molecular weight: 124 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt (Intra) Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

追加情報

Is unsuitable for ChIP.

Target information above from: UniProt accession Q9H2P0 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: 23394 Human
- Entrez Gene: 11538 Mouse
- Entrez Gene: 64622 Rat
- Omim: 611386 Human
- SwissProt: Q9H2P0 Human
- SwissProt: Q9Z103 Mouse
- SwissProt: Q9JKL8 Rat
- Unigene: 570355 Human
- Unigene: 201322 Mouse
- Unigene: 48777 Rat
see all
別名
- Activity dependent neuroprotective protein antibody
- Activity dependent neuroprotector antibody
- Activity-dependent neuroprotective protein antibody
- Activity-dependent neuroprotector homeobox protein antibody
- Adnp antibody
- ADNP_HUMAN antibody
- KIAA0784 antibody
see all

Western blot - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

All lanes : Anti-ADNP antibody [EPR25434-4] (ab300114) at 1/1000 dilution

Lane 1 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 124 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

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

Exposure time:

Lanes 1-3: 114 seconds; Lane 4: 59 seconds.

Lysates should be made freshly and used in WB immediately to minimize protein degradation.
Western blot - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

All lanes : Anti-ADNP antibody [EPR25434-4] (ab300114) at 1/1000 dilution

Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 2 : U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 3 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 124 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

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

Lysates should be made freshly and used in WB immediately to minimize protein degradation
Western blot - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

All lanes : Anti-ADNP antibody [EPR25434-4] (ab300114) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 124 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

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

Lysates should be made freshly and used in WB immediately to minimize protein degradation. We used fresh mouse brain tissue lysate.

This blot was developed using a higher sensitivity ECL substrate.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labelling ADNP with ab300114 at 1/100 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on human colon carcinoma is observed (PMID: 27903678). The section was incubated with ab300114 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-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling ADNP with ab300114 at 1/100 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Nuclear staining on mouse cerebrum is observed. The section was incubated with ab300114 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-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse glioblastoma tissue labelling ADNP with ab300114 at 1/100 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on mouse glioblastoma is observed. The section was incubated with ab300114 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-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labelling ADNP with ab300114 at 1/100 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on rat cerebrum is observed. The section was incubated with ab300114 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-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum (fresh) tissue labeling ADNP with ab300114 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: PBS was used instead of ab300114 followed by preadsorbed secondary antibody (ab150081) Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Immunohistochemistry (Frozen sections) - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum (fresh) tissue labeling ADNP with ab300114 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (Blue).

PBS was used instead of ab300114 followed by preabsorbed secondary antibody (ab150081) Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (ab300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized HeLa (human cervix adenocarcinoma epithelial cell) cells lebelling ADNP with ab300114 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line. Tubuline was stained with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: PBS was used instead of ab300114 followed by preabsorbed secondary antibody (ab150081) Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (ab300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized NIH/3T3 (mouse embryonic fibroblast) cells lebelling ADNP with ab300114 at 1/50 (11.22 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2µg/mL) dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody control: PBS was used instead of ab300114 followed by preabsorbed secondary antibody (ab150081) Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 µg/mL) dilution.
Flow Cytometry (Intracellular) - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed and 90% methnol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling ADNP with ab300114 at 1/500 dilution (Red) compared with a rabbit monoclonal IgG (ab172730) (Black) isotype control. Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling ADNP with ab300114 at 1/500 dilution (Red) (Red) compared with a rabbit monoclonal IgG (ab172730) (Black) isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

ADNP was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab300114 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab300114 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 (Input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg

Lane 2 (+): ab300114 IP in HeLa whole cell lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab300114 in HeLa whole cell lysate

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

Exposure time: 84 seconds
Immunoprecipitation - Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (AB300115)

This data was developed using ab300114, the same antibody clone in a different buffer formulation.

ADNP was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab300114 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab300114 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 (Inset): NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 µg

Lane 2 (+): ab300114 IP in NIH/3T3 whole cell lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab300114 in NIH/3T3 whole cell lysate

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

Exposure time: 3 minutes.
Anti-ADNP antibody [EPR25434-4] (BSA and Azide free) (ab300115)

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

Datasheet download

Download

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ab300115 は論文での使用が確認できていません。

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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

キャリア・フリー

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

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

Datasheet download

Certificate of Compliance

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