Anti-Connexin 32 / GJB1 抗体 [EPR26417-44] (ab300401)

製品名

Anti-Connexin 32 / GJB1 antibody [EPR26417-44]
Connexin 32 / GJB1 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR26417-44] to Connexin 32 / GJB1
由来種

Rabbit
特異性

This antibody may not be suitable for cell lines in Western Blot.
アプリケーション

適用あり: WB, IHC-P, 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

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: Mouse liver, mouse spinal cord, rat liver, rat spinal cord, human spinal cord. IHC-P: Human liver, human spinal cord, mouse liver, mouse brain stem, rat liver, and HEK-293T transfected with a mouse Gjb1 expression vector tissues. IHC-Fr: Mouse liver, rat liver tissues. IP: Mouse liver cell.
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

EPR26417-44
アイソタイプ

IgG
研究分野
- Neuroscience
- Processes

Alternative Versions
- Anti-Connexin 32 / GJB1 antibody [EPR26417-44] - BSA and Azide free (ab300413)
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ab300401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		1/1000.
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr		1/100.
IP		1/30.

特記事項
WB 1/1000.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/100.
IP 1/30.

追加情報

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

機能

One gap junction consists of a cluster of closely packed pairs of transmembrane channels, the connexons, through which materials of low MW diffuse from one cell to a neighboring cell.
関連疾患

Defects in GJB1 are the cause of Charcot-Marie-Tooth disease X-linked type 1 (CMTX1) [MIM:302800]; also designated CMT-X. CMTX1 is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathies characterized by severely reduced motor nerve conduction velocities (NCVs) (less than 38m/s) and segmental demyelination and remyelination, and primary peripheral axonal neuropathies characterized by normal or mildly reduced NCVs and chronic axonal degeneration and regeneration on nerve biopsy. CMTX1 has both demyelinating and axonal features. Central nervous system involvement may occur.
Defects in GJB1 may contribute to the phenotype of Dejerine-Sottas syndrome (DSS) [MIM:145900]; also known as Dejerine-Sottas neuropathy (DSN) or hereditary motor and sensory neuropathy III (HMSN3). DSS is a severe degenerating neuropathy of the demyelinating Charcot-Marie-Tooth disease category, with onset by age 2 years. DSS is characterized by motor and sensory neuropathy with very slow nerve conduction velocities, increased cerebrospinal fluid protein concentrations, hypertrophic nerve changes, delayed age of walking as well as areflexia. There are both autosomal dominant and autosomal recessive forms of Dejerine-Sottas syndrome.
配列類似性

Belongs to the connexin family. Beta-type (group I) subfamily.
細胞内局在

Cell membrane. Cell junction > gap junction.
Target information above from: UniProt accession P08034 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: 2705 Human
- Entrez Gene: 14618 Mouse
- Entrez Gene: 29584 Rat
- Omim: 304040 Human
- SwissProt: P08034 Human
- SwissProt: P28230 Mouse
- SwissProt: P08033 Rat
- Unigene: 333303 Human
- Unigene: 21198 Mouse
- Unigene: 10444 Rat
see all
別名
- Charcot Marie Tooth neuropathy X linked antibody
- CMTX 1 antibody
- CMTX antibody
- CMTX1 antibody
- Connexin-32 antibody
- Connexin32 antibody
- CX 32 antibody
- Cx32 antibody
- CXB1_HUMAN antibody
- GAP junction 28 kDa liver protein antibody
- Gap junction beta 1 protein antibody
- Gap junction beta-1 protein antibody
- Gap junction protein beta 1 32kD antibody
- Gap junction protein beta 1 antibody
- GJB 1 antibody
- GJB1 antibody
see all

Western blot - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

All lanes : Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (ab300401) at 1/1000 dilution

Lane 1 : Mouse liver tissue lysate
Lane 2 : Mouse spinal cord tissue lysate
Lane 3 : Mouse spleen tissue lysate
Lane 4 : Mouse lung tissue lysate
Lane 5 : Rat liver tissue lysate
Lane 6 : Rat spinal cord tissue lysate
Lane 7 : Rat spleen tissue lysate

Lysates/proteins at 20 µg per lane.

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

Observed band size: 26, 48 kDa why is the actual band size different from the predicted?

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

26-kDa Cx32 monomers and 48-kDa Cx32 dimers are observed. The molecular weight observed is consistent with what has been described in the literature (PMID: 17972320).

A band around 40 kDa may due to degradation.

Low expression tissue: spleen, lung(PMID: 2557354).

Exposure time: Lanes 1-4: 3 minutes; Lanes 5-7: 59 seconds
Western blot - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

All lanes : Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (ab300401) at 1/1000 dilution

Lane 1 : Human spinal cord tissue lysate
Lane 2 : Human lung tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
Lane 1 : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution
Lane 2 : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 26, 48 kDa why is the actual band size different from the predicted?

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

26-kDa Cx32 monomers and 48-kDa Cx32 dimers are observed. The molecular weight observed is consistent with what has been described in the literature (PMID: 17972320).

Low expression tissue: lung (PMID: 2557354).

Exposure time: 125 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of paraffin-embedded human liver tissue labelling Connexin 32/ GJB1 with ab300401 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining in human liver. The section was incubated with ab300401 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 a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of paraffin-embedded human spinal cord tissue labelling Connexin 32 / GJB1 with ab300401 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining in human spinal cord. The section was incubated with ab300401 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 a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labelling Connexin 32 / GJB1 with ab300401 at 1/10000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining in mouse liver. The section was incubated with ab300401 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 a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of paraffin-embedded mouse brain stem tissue labelling Connexin 32 / GJB1 with ab300401 at 1/10000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining in mouse brain stem. The section was incubated with ab300401 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 a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labelling Connexin 32 / GJB1 with ab300401 at 1/10000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining in rat liver. The section was incubated with ab300401 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 a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of paraffin-embedded HEK-293T tissue labelling Connexin 32 / GJB1 with ab300401 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining on (A) HEK-293T transfected with a mouse Gjb1 expression vector, no staining on (B) HEK-293T transfected with empty vector. The section was incubated with ab300401 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 a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue labelling Connexin 32 / GJB1 with ab300401 at 1/10000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Negative control: no staining in mouse cardiac muscle. The section was incubated with ab300401 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 a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Frozen sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labeling Connexin 32 / GJB1 with ab300401 at 1/100 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Positive staining on mouse liver is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.
Immunohistochemistry (Frozen sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen (fresh) tissue labeling Connexin 32 / GJB1 with ab300401 at 1/100 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Low expression control (PMID: 2557354). No staining on mouse spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: PBS was used instead of primary antibody followed by secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.
Immunohistochemistry (Frozen sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labeling Connexin 32 / GJB1 with ab300401 at 1/100 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Positive staining on rat liver is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: PBS was used instead of primary antibody followed by secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.
Immunohistochemistry (Frozen sections) - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat spleen (fresh) tissue labeling Connexin 32 / GJB1 with ab300401 at 1/100 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Low expression control (PMID: 2557354).No staining on rat spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.
Immunoprecipitation - Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

Connexin 32 / GJB1 was immunoprecipitated from 0.35 mg mouse liver tissue lysate 10 μg with ab300401 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300401 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: mouse liver tissue lysate 10 μg

Lane 2: ab300401 IP in mouse liver tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab260035 in mouse liver tissue lysate

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

Exposure time: 6 seconds

26-kDa Cx32 monomer, 48-kDa Cx32 dimers and 75-kDa trimers are observed. The molecular weight observed is consistent with what has been described in the literature (PMID: 17972320).
Anti-Connexin 32 / GJB1 antibody [EPR26417-44] (AB300401)

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

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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

Isotype control

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

関連疾患

配列類似性

細胞内局在

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

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