Name: Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5]
SKU: ab110333
Rating: 4 (6 reviews)

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Western blot - Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333)

Key features and details

Mouse monoclonal [13G2AE2BH5] to PDHX
Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Cow, Human
Isotype: IgG2a

製品名

Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5]
PDHX 一次抗体製品一覧
製品の詳細

Mouse monoclonal [13G2AE2BH5] to PDHX
由来種

Mouse
アプリケーション

適用あり: WB, ICC/IF, Flow Cyt, IHC-Pmore details
種交差性

交差種: Mouse, Rat, Cow, Human
免疫原

Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- Isolated mitochondria from Human, Bovine, Rat and Mouse heart and HepG2 lysate; cultured, normal Human embryonic lung fibroblasts (strain MRC5); Human cerebellum tissue; HL60 cells.
特記事項

This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Product was previously marketed under the MitoSciences sub-brand.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C. Do Not Freeze.
バッファー

pH: 7.5
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline
Concentration information loading...
精製度

IgG fraction
特記事項（精製）

ab110333 was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
ポリ/モノ

モノクローナル
クローン名

13G2AE2BH5
アイソタイプ

IgG2a
軽鎖の種類

kappa
研究分野

Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
- Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)

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ab110333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	(4)	Use a concentration of 1 µg/ml. Predicted molecular weight: 54 , 69 kDa.
ICC/IF	(2)	Use a concentration of 1 µg/ml. (heat-induced antigen-retrieval improves signal)
Flow Cyt		Use a concentration of 1 µg/ml. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
IHC-P		1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

特記事項
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 54 , 69 kDa.
ICC/IF Use a concentration of 1 µg/ml. (heat-induced antigen-retrieval improves signal)
Flow Cyt Use a concentration of 1 µg/ml. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
IHC-P 1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

機能

Required for anchoring dihydrolipoamide dehydrogenase (E3) to the dihydrolipoamide transacetylase (E2) core of the pyruvate dehydrogenase complexes of eukaryotes. This specific binding is essential for a functional PDH complex.
関連疾患

Defects in PDHX are the cause of pyruvate dehydrogenase E3-binding protein deficiency (PDHXD) [MIM:245349].
配列類似性

Belongs to the 2-oxoacid dehydrogenase family.
Contains 1 lipoyl-binding domain.
細胞内局在

Mitochondrion matrix.
Target information above from: UniProt accession O00330 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: 8050 Human
- Entrez Gene: 27402 Mouse
- Entrez Gene: 311254 Rat
- Omim: 608769 Human
- SwissProt: O00330 Human
- SwissProt: Q8BKZ9 Mouse
- Unigene: 502315 Human
- Unigene: 315011 Mouse
別名
- Dihydrolipoamide dehydrogenase binding protein of pyruvate dehydrogenase complex antibody
- Dihydrolipoamide dehydrogenase-binding protein of pyruvate dehydrogenase complex antibody
- DLDBP antibody
- E3 binding protein antibody
- E3-binding protein antibody
- E3BP antibody
- Lipoyl containing pyruvate dehydrogenase complex component X antibody
- Lipoyl-containing pyruvate dehydrogenase complex component X antibody
- mitochondrial antibody
- ODPX_HUMAN antibody
- OPDX antibody
- PDHX antibody
- PDX 1 antibody
- PDX1 antibody
- Pro X antibody
- proX antibody
- Pyruvate dehydrogenase complex component X antibody
- Pyruvate dehydrogenase complex lipoyl containing component X antibody
- Pyruvate dehydrogenase complex, E3 binding protein subunit antibody
- Pyruvate dehydrogenase protein X component antibody
- Pyruvate dehydrogenase protein X component mitochondrial antibody
see all

Western blot - Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333)

All lanes : Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333) at 1 µg/ml

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : DLAT knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HL-60 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 54 , 69 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab110333 observed at 72 kDa. Red - loading control, ab181602, observed at 38 kDa.

ab110333 was shown to specifically react with in wild-type HAP1 cells as signal was lost in DLAT knockout cells. Wild-type and DLAT knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab110333 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333)

All lanes : Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333) at 1 µg/ml

Lane 1 : Isolated mitochondria from Human heart at 5 µg
Lane 2 : Isolated mitochondria from Bovine heart at 1 µg
Lane 3 : Isolated mitochondria from Rat heart at 10 µg
Lane 4 : Isolated mitochondria from Mouse heart at 10 µg
Lane 5 : HepG2 cell lysate at 20 µg

Predicted band size: 54 , 69 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333)This image is courtesy of an Abreview submitted by Dimitra Kalamida

ab110333 staining Pyruvate dehydrogenase E2/E3bp in Human HUVEC by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 pH 7.4 for 5 minutes and blocked with 5% BSA for 20 minutes at room temperature. Samples were incubated with primary antibody (1/1000 in PBS) for 1 hour. A CF568-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333)

Immunocytochemistry analysis using ab110333 at 1µg/ml staining Pyruvate dehydrogenase E2/E3bp in cultured, normal Human embryonic lung fibroblasts and an AlexaFluor® 488 goat anti-mouse IgG2a secondary antibody (2 ug/ml).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333)

Immunohistological analysis using ab110333 at 1/1000 dilution staining Pyruvate dehydrogenase E2/E3bp in Human cerebellum tissue (Formalin-fixed, Paraffin-embedded).
Note: Immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells.
Flow Cytometry - Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333)

Flow cytometric analysis using ab110333 at 1µg/ml staining Pyruvate dehydrogenase E2/E3b in HL60 cells (blue). Isotype control antibody (red).

Mouse on Mouse staining protocol

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

ab110333 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。

ab110333 は 46 報の論文で使用されています。

Allen FM et al. Rapid fractionation of mitochondria from mouse liver and heart reveals in vivo metabolite compartmentation. FEBS Lett 597:246-261 (2023). PubMed: 36217875
Yamada T et al. Prevention and regression of megamitochondria and steatosis by blocking mitochondrial fusion in the liver. iScience 25:103996 (2022). PubMed: 35310936
Salvany S et al. Accumulation of misfolded SOD1 outlines distinct patterns of motor neuron pathology and death during disease progression in a SOD1G93A mouse model of amyotrophic lateral sclerosis. Brain Pathol 32:e13078 (2022). PubMed: 35584812
Küey C et al. Recruitment of clathrin to intracellular membranes is sufficient for vesicle formation. Elife 11:N/A (2022). PubMed: 35852853
Ramirez A et al. Cannabidiol activates PINK1-Parkin-dependent mitophagy and mitochondrial-derived vesicles. Eur J Cell Biol 101:151185 (2022). PubMed: 34915361

View all Publications for this product

レビューと Q&A

Show All レビュー Q&A

レビューを送る質問を送る

1-8 of 8 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (HEK293)

Gel Running Conditions

Reduced Denaturing (4-20%)

Loading amount

10 µg

Specification

HEK293

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jun 30 2022

Application

Western blot

Sample

Human Cell lysate - whole cell (Dopaminergic Neurons)

Gel Running Conditions

Reduced Denaturing (NuPage 4-12% MES buffer)

Loading amount

30 µg

Specification

Dopaminergic Neurons

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Oct 22 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (colon cancer, kidney)

Gel Running Conditions

Reduced Denaturing

Loading amount

30 µg

Specification

colon cancer, kidney

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Oct 16 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Rat Cell (H9C2)

Permeabilization

Yes - 0.1% Triton in PBS for 10 min

Specification

H9C2

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 23°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Nov 17 2017

Application

Western blot

Sample

Human Cell lysate - whole cell (prostate cancer cell line)

Gel Running Conditions

Reduced Denaturing (10% acrylamide gel)

Loading amount

20 µg

Specification

prostate cancer cell line

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Stamatia Pouliliou

Verified customer

投稿 Jun 26 2015

Application

Immunocytochemistry/ Immunofluorescence

Blocking step

BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C

Sample

Human Cell (HuVEC)

Specification

HuVEC

Permeabilization

Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Dimitra Kalamida

Verified customer

投稿 Nov 18 2013

Question

Thank you for your assistance. I really appreciate it.
I'm still wondering regarding the ab109902. What is the benefit of using pyruvate dehydrogenase E2/E3 as immonogen for the antibody? Why not E1 subunit?

Abcam community

Verified customer

Asked on Oct 18 2012

Answer

Thank you for your reply.

The kit ab109902 measures the activity of the whole PDH complex by assaying generated NADH. This requires all three activities (pyruvate dehydrogenase –E1, dihydrolipoyl acyltransferase-E2 and dihydrolipoyl dehydrogenase-E3) thus the purpose of the capture antibody (such as ab110333) is to capture whole PDH complex without interfering with the activity. We tested number of capture antibodies against different subunits but ab110333 showed to be optimal. Obviously, another important step in the protocol is to solubilize the biological material without dissociating the subunits of the PDH complex. The immunogen used to generate ab110333 was whole PDH complex, the target of the antibody was determined subsequently.

I hope this information helps. Please contact us with any other questions.

Abcam Scientific Support

Answered on Oct 18 2012

Question

Inquiry: Hi, I am interested in trying this product with rat samples. Do you have any direct evidence for its efficacy against rat PDH, do you know if it was raised against a sequence that is identical to rat? The product information states that it reacts with human and cow, and I am concerned that there are no "predicted" interactions despite the high sequence identity of PDH subunits between human, mouse and rat, etc.

Abcam community

Verified customer

Asked on Apr 24 2012

Answer

Thank you for contacting us.

I have had the opportunity to speak with the lab regarding this on your behalf. They recommend using ab110333 (MSP06) with rat samples. The antibody targets on E2/E3 binding protein but pulls down the whole PDH complex using IP reagents and protocol. The product page for ab110333 is located at:

https://www.abcam.com/Pyruvate-dehydrogenase-E2-E3bp-antibody-13G2AE2BH5-ab110333.html

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Abcam Scientific Support

Answered on Apr 24 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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Key features and details

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

特記事項（精製）

ポリ/モノ

クローン名

アイソタイプ

軽鎖の種類

研究分野

関連製品

Compatible Secondaries

Conjugation kits

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

参考文献 (46)

レビューと Q&A

Western blot abreview for Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5]

Western blot abreview for Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5]

Western blot abreview for Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5]

Western blot abreview for Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5 ]