CISD1/MitoNEET重组抗体[EPR29116-27]_CISD1/MitoNEET抗体(ab318200)

产品名称

Anti-CISD1/MitoNEET抗体[EPR29116-27] - BSA and Azide free
参阅全部 CISD1/MitoNEET 一抗
描述

兔单克隆抗体[EPR29116-27] to CISD1/MitoNEET - BSA and Azide free
宿主

Rabbit
特异性

This antibody weakly cross-react with human CISD2 overexpression lysate.
经测试应用

适用于: IHC-P, ICC/IF, WBmore details
不适用于: Flow Cyt (Intra) or IP
种属反应性

与反应: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: Wild-type HeLa,293T, Neuro-2a, NIH/3T3, HeLa mitochondrial fraction, Human cerebellum, Mouse cerebellum, Mouse liver, Mouse kidney, Rat cerebellum, Rat liver and C6 lysates. IHC-P: Human cerebrum, Human colon, Human ovarian carcinoma, Mouse liver, Mouse colon, Mouse liver cancer, Rat liver, Rat colon and Wild-type Hela tissues. ICC/IF: Wildtype HeLa and NIH/3T3 cells.
常规说明
ab318200 is the carrier-free version of ab318199
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.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C.
存储溶液

pH: 7.2
Constituent: 100% PBS
无载体

是
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR29116-27
同种型

IgG

Alternative Versions
- Anti-CISD1/MitoNEET antibody [EPR29116-27] (ab318199)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120)
KO cell lines
- Human CISD1 (MitoNEET) knockout HeLa cell line (ab265276)
KO cell lysates
- Human CISD1 (MitoNEET) knockout HeLa cell lysate (ab258817)
Related Products
- Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
- Anti-AIF antibody [E20] - Mitochondrial Marker (ab32516)

The Abpromise guarantee

Abpromise™承诺保证使用ab318200于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
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.
ICC/IF		Use at an assay dependent concentration.
WB		Use at an assay dependent concentration. Predicted molecular weight: 12 kDa.

说明
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.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 12 kDa.

应用说明

Is unsuitable for Flow Cyt (Intra) or IP.

功能

Plays a key role in regulating maximal capacity for electron transport and oxidative phosphorylation (By similarity). May be involved in Fe-S cluster shuttling and/or in redox reactions.
组织特异性

Expression is reduced in cells derived from cystic fibrosis patients.
序列相似性

Belongs to the CISD protein family.
翻译后修饰

Ubiquitinated by PARK2 during mitophagy, leading to its degradation and enhancement of mitophagy. Deubiquitinated by USP30.
细胞定位

Mitochondrion outer membrane.
Target information above from: UniProt accession Q9NZ45 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 55847 Human
- SwissProt: Q9NZ45 Human
- Unigene: 370102 Human
别名
- C10orf70 antibody
- CDGSH iron-sulfur domain-containing protein 1 antibody
- CISD1 antibody
- CISD1_HUMAN antibody
- MitoNEET antibody
- ZCD1 antibody
see all

Western blot - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

All lanes : Anti-CISD1/MitoNEET antibody [EPR29116-27] (ab318199) at 1/1000 dilution

Lane 1 : Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : CISD1 knockout HeLa whole cell lysate
Lane 3 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
Lane 5 : NIH/3T3 (mouse embryonic fibroblast) whole cell 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

Performed under reducing conditions.

Predicted band size: 12 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

This data was developed using ab318199, the same antibody clone in a different buffer formulation.

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

In Western blot, ab318199 was shown to bind specifically to CISD1. Target of interest was observed at 15 kDa in wild-type Hela cell lysates (lane 1) with no signal observed at this size in CISD1 knockout cell line (lane 2) (lane 2, knockout cell line ab265276 / knockout cell lysate ab258817).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time: Lane 1-2: 114 seconds, Lane 3-5: 59 seconds
Western blot - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

All lanes : Anti-CISD1/MitoNEET antibody [EPR29116-27] (ab318199) at 1/1000 dilution

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) cytoplasmic fraction
Lane 2 : HeLa mitochondrial fraction

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: 12 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

This data was developed using ab318199, the same antibody clone in a different buffer formulation.

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-AIF antibody [E20] (ab32516) staining at 1/1000 dilution.
Western blot - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

All lanes : Anti-CISD1/MitoNEET antibody [EPR29116-27] (ab318199) at 1/1000 dilution

Lane 1 : Human cerebellum tissue lysate
Lane 2 : Mouse cerebellum tissue lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : Mouse kidney tissue lysate
Lane 5 : Rat cerebellum tissue lysate
Lane 6 : Rat liver tissue lysate
Lane 7 : C6 (rat glial tumor glial cell) whole cell 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: 12 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

Exposure time: 8 seconds

This data was developed using ab318199, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling MitoNEET with ab318199 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on human cerebrum. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling MitoNEET with ab318199 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on human colon. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling MitoNEET with ab318199 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on human ovarian carcinoma. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling MitoNEET with ab318199 at 1/5000 (0.102 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on mouse liver. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling MitoNEET with ab318199 at 1/5000 (0.102 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on mouse colon. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse liver cancer tissue labeling MitoNEET with ab318199 at 1/5000 (0.102 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on mouse liver cancer. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling MitoNEET with ab318199 at 1/5000 (0.102 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on rat liver. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling MitoNEET with ab318199 at 1/5000 (0.102 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on rat colon. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded A Wild-type Hela (human cervical adenocarcinoma epithelial cell) cell pellet B CISD1 knockout Hela cell pellet tissue labeling MitoNEET with ab318199 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Positive staining on (A) wild-type Hela cell pellet, no staining on (B) human CISD1 knockout Hela cell pellet. The section was incubated with ab318199 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CISD1 KO HeLa (human cervical adenocarcinoma epithelial cell), ab265276 cells labelling MitoNEET with ab318199 at 1/100 (5.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing mitochondrial staining in wildtype HeLa cells (shown in green), and negative staining in CISD1 knockout HeLa cells. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/1000 1ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1: ab318199 at 1/100 (5.1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2 ug/mL dilution.
-ve control 2: Anti-COX IV mouse monoclonal 1/1000 1ug/ml dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Immunocytochemistry/ Immunofluorescence - Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

This data was developed using ab318199, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling MitoNEET with ab318199 at 1/100 (5.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing mitochondrial staining in NIH/3T3 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/1000 1ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1: ab318199 at 1/100 (5.1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2 ug/mL dilution.
-ve control 2: Anti-COX IV mouse monoclonal 1/1000 1ug/ml dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Anti-CISD1/MitoNEET antibody [EPR29116-27] - BSA and Azide free (ab318200)

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

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概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

无载体

纯度

克隆

克隆编号

同种型

相关产品

Alternative Versions

Compatible Secondaries

KO cell lines

KO cell lysates

Related Products

应用

The Abpromise guarantee

靶标

功能

组织特异性

序列相似性

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

Datasheet download

Certificate of Compliance

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