使用敲除细胞株进行验证重组RabMAb

重组Anti-Acetyl Coenzyme A Carboxylase抗体[EP687Y] (ab45174)

Reviews (3)Q&A (3)References (23)
Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
  • Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
  • Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
  • Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP687Y] to Acetyl Coenzyme A Carboxylase
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

概述

  • 产品名称

    Anti-Acetyl Coenzyme A Carboxylase抗体[EP687Y]
    参阅全部 Acetyl Coenzyme A Carboxylase 一抗

  • 描述

    兔单克隆抗体[EP687Y] to Acetyl Coenzyme A Carboxylase

  • 宿主

    Rabbit

  • 特异性

    Acetyl Coenzyme A Carboxylase is highly expressed in lipogenic tissues such as liver, adipose, and lactating mammary gland, and its activities are regulated at various levels [Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7515-20.].

  • Tested Applications & Species

    Application Species
    IHC-P
    Mouse
    Rat
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • 免疫原

    Synthetic peptide within Human Acetyl Coenzyme A Carboxylase aa 350-450. The exact sequence is proprietary.
    (Peptide available as ab195232)

  • 阳性对照

    • WB: HAP1, HeLa, C2C12 and A431 cell lysate; Rat adrenal gland tissue lysates IHC-P: Human liver and breast carcinoma tissue, Rat stomcah tissue, Mouse kidney tissue

  • 常规说明

     

     

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

应用

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab45174 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
IHC-P
Mouse
Rat
Human
WB
Mouse
Rat
Human
应用 Ab评论 说明
WB (2)
1/1000 - 1/2000. Detects a band of approximately 265 kDa (predicted molecular weight: 265 kDa).Can be blocked with Acetyl Coenzyme A Carboxylase peptide (ab195232).
IHC-P
1/400. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
说明
WB
1/1000 - 1/2000. Detects a band of approximately 265 kDa (predicted molecular weight: 265 kDa).Can be blocked with Acetyl Coenzyme A Carboxylase peptide (ab195232).
IHC-P
1/400. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

靶标

  • 功能

    Catalyzes the rate-limiting reaction in the biogenesis of long-chain fatty acids. Carries out three functions: biotin carboxyl carrier protein, biotin carboxylase and carboxyltransferase.

  • 组织特异性

    Expressed in brain, placental, skeletal muscle, renal, pancreatic and adipose tissues; expressed at low level in pulmonary tissue; not detected in the liver.

  • 通路

    Lipid metabolism; malonyl-CoA biosynthesis; malonyl-CoA from acetyl-CoA: step 1/1.

  • 疾病相关

    Acetyl-CoA carboxylase 1 deficiency

  • 序列相似性

    Contains 1 ATP-grasp domain.
    Contains 1 biotin carboxylation domain.
    Contains 1 biotinyl-binding domain.
    Contains 1 carboxyltransferase domain.

  • 翻译后修饰

    Phosphorylation on Ser-1263 is required for interaction with BRCA1.

  • 细胞定位

    Cytoplasm.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • ACAC antibody
    • ACACA antibody
    • ACACA_HUMAN antibody
    • ACACB antibody
    • ACC alpha antibody
    • ACC antibody
    • ACC beta antibody
    • ACC-alpha antibody
    • ACC1 antibody
    • ACC2 antibody
    • ACCA antibody
    • ACCB antibody
    • Acetyl CoA carboxylase 1 antibody
    • Acetyl CoA carboxylase 2 antibody
    • Acetyl CoA carboxylase alpha antibody
    • Acetyl CoA carboxylase beta antibody
    • Acetyl Coenzyme A carboxylase alpha antibody
    • Acetyl Coenzyme A carboxylase beta antibody
    • Biotin carboxylase antibody
    • COA1 antibody
    • COA2 antibody
    • HACC275 antibody
    • OTTHUMP00000164069 antibody
    • OTTHUMP00000164070 antibody
    • OTTHUMP00000164076 antibody
    • OTTHUMP00000240532 antibody
    see all

图片

  • Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    All lanes : Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/2000 dilution (unpurified)

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : ACACA (Acetyl Coenzyme A Carboxylase) knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : A431 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 265 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab45174 observed at 265 kDa. Red - loading control, ab130007, observed at 130 kDa.

    ab45174 was shown to specifically react with Acetyl Coenzyme A Carboxylase in wild-type HAP1 cells as signal was lost in ACACA (Acetyl Coenzyme A Carboxylase) knockout cells. Wild-type and ACACA (Acetyl Coenzyme A Carboxylase) knockout samples were subjected to SDS-PAGE. Ab45174 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Acetyl Coenzyme A Carboxylase with purified ab45174 at 1/400 dilution (2.06 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    All lanes : Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/1000 dilution (Purified)

    Lane 1 : C2C12 (Mouse myoblasts myoblast) whole cell lysates
    Lane 2 : Rat adrenal gland lysates

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 265 kDa
    Observed band size: 265 kDa



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

    We are unsure how to define the extra bands.

  • Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/1000 dilution (Purified) + A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 15 µg

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

    Predicted band size: 265 kDa
    Observed band size: 265 kDa



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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomcah tissue sections labeling Acetyl Coenzyme A Carboxylase with purified ab45174 at 1/400 dilution (2.06 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling Acetyl Coenzyme A Carboxylase with purified ab45174 at 1/400 dilution (2.06 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)

    ab45174 (unpurified), at a dilution of 1/50, staining human Acetyl Coenzyme A Carboxylase in human liver by immunohistochemistry using paraffin embedded tissue. Heat mediated antigen retrieval was perfromed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)

    Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Acetyl Coenzyme A Carboxylase with ab45174 (unpurified) at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)
    Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174)

实验方案

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

文献 (23)

发表研究结果有使用 ab45174？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab45174 被引用在 23 文献中.

  • Li M  et al. Sirt3 modulates fatty acid oxidation and attenuates cisplatin-induced AKI in mice. J Cell Mol Med 24:5109-5121 (2020). PubMed: 32281286
  • Zhou Y  et al. PMP22 Regulates Cholesterol Trafficking and ABCA1-Mediated Cholesterol Efflux. J Neurosci 39:5404-5418 (2019). PubMed: 31061090
  • Chen XY  et al. LB100 ameliorates nonalcoholic fatty liver disease via the AMPK/Sirt1 pathway. World J Gastroenterol 25:6607-6618 (2019). PubMed: 31832001
  • Jia H  et al. Perilipin 5 promotes hepatic steatosis in dairy cows through increasing lipid synthesis and decreasing very low density lipoprotein assembly. J Dairy Sci 102:833-845 (2019). PubMed: 30415861
  • Chen Y  et al. Long noncoding RNA FLRL2 alleviated nonalcoholic fatty liver disease through Arntl-Sirt1 pathway. FASEB J 33:11411-11419 (2019). PubMed: 31311301
View all Publications for this product

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1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
50 µg
Specification
Liver
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

提交于 Jul 02 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y]

 Average
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse hepatoma cell line)
Permeabilization
Yes - 0.5% Triton X-100
Specification
mouse hepatoma cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

提交于 Aug 08 2019

Western blot abreview for Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (C2C12 differentiated myotube)
Gel Running Conditions
Reduced Denaturing (4-12% Nupage Tris-Bis gel)
Loading amount
10 µg
Specification
C2C12 differentiated myotube
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More

Abcam user community

Verified customer

提交于 Aug 28 2017

Question

In product datasheet, Acetyl Coenzyme A Carboxylase antibody(ab45174) is made on human sequence,the epitope region is at 392-407a.a., and the band size is 265kDa (ACC1)in western blot. I searched NCBI for ACC1 homo sapience and found 5 isoforms of ACC1(NP_942131-NP_942136). Is the epitope region at 392-407a.a. of ACC1 isoform1(NP_942131)? I want to know the epitope sequence.

Read More
Answer

Thank you for your enquiry. I have contacted the originator of the product for the information you required. They have confirmed that the epitope region is indeed at 392-407 a.a. I hope this information will be helpful to you. If there is anything else that I can help you with, please do not hesitate to contact me.

Read More

Question

Can you tell me if this antibody will react with ACC2? What is the animal species that the immunogen was made from? Also, please where is the epitope region?

Read More
Answer

Thank you for your enquiry. I can confirm that the band in the WB image is the ACC1 band. There is a 75% homology between ACC1 and ACC2. Generally, we say that if they share 85% or more, they will cross react, so I don’t think these will, but I cannot say for sure. I am afraid that the we do not have the animal species information. However, the peptides are synthetically made based on human sequence. The peptide is 15 a.a. long and the epitope region is at 392~407 a.a.

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Question

I am looking for a good quality antibody against ACC, in particular one which will recognize ACC from human and mouse origin. First, from the western blot on the data sheet, this antibody seems to be very sensitive. Does this antibody recognize both ACC1 and ACC2? Has this antibody been used to detect ACC protein from tissues? Thank you for your help

Read More
Answer

Thank you for your enquiry. An alignment of the peptide used to make ab45174 with human ACC2 (http://www.expasy.org/uniprot/COA2_HUMAN) shows 75% homology so there is a chance that ab45174 will cross react with ACC2. The antibody has only been tested in WB assays using A431 cell lysates. If you have any other questions, please do not hesitate to contact us.

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