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使用敲除细胞株进行验证

Anti-CPT1A抗体[8F6AE9] (ab128568)

Reviews (5)Q&A (3)References (179)
Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)
  • Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [8F6AE9] (ab128568)
  • Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [8F6AE9] (ab128568)
  • Flow Cytometry - Anti-CPT1A antibody [8F6AE9] (ab128568)
  • Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)
  • In-Cell ELISA - Anti-CPT1A antibody [8F6AE9] (ab128568)

Key features and details

  • Mouse monoclonal [8F6AE9] to CPT1A
  • Suitable for: Flow Cyt, IHC-P, WB, ICC/IF, In-Cell ELISA
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG2b

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 HRP

概述

  • 产品名称

    Anti-CPT1A抗体[8F6AE9]
    参阅全部 CPT1A 一抗

  • 描述

    小鼠单克隆抗体[8F6AE9] to CPT1A

  • 宿主

    Mouse

  • 经测试应用

    适用于: Flow Cyt, IHC-P, WB, ICC/IF, In-Cell ELISAmore details

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Recombinant fragment, corresponding to amino acids 489-773 of Human CPT1A.

  • 阳性对照

    • Partial Human Recombinant CPT1A protein; HepG2, H9C2, H4IIE Whole Cell Lysates; Rat heart mitochondrial, Human liver, Rat liver and Mouse liver homogenates; H9C2 cells; HeLa cells IHC-P: human normal kidney FFPE tissue sections

  • 常规说明

    This monoclonal antibody to CPT1A has been knockout validated in Western blot. The expected band for CPT1A was observed in wild type cells and the band was not seen in CPT1A knockout cells.

    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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab128568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
Flow Cyt
Use at an assay dependent concentration.
IHC-P (1)
Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB (3)
Use a concentration of 1 µg/ml. Predicted molecular weight: 88 kDa.
ICC/IF (1)
Use a concentration of 0.5 - 5 µg/ml.
In-Cell ELISA
Use a concentration of 0.1 - 1 µg/ml.
说明
Flow Cyt
Use at an assay dependent concentration.
IHC-P
Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 88 kDa.
ICC/IF
Use a concentration of 0.5 - 5 µg/ml.
In-Cell ELISA
Use a concentration of 0.1 - 1 µg/ml.

靶标

  • 组织特异性

    Strong expression in kidney and heart, and lower in liver and skeletal muscle.

  • 通路

    Lipid metabolism; fatty acid beta-oxidation.

  • 疾病相关

    Defects in CPT1A are the cause of carnitine palmitoyltransferase 1A deficiency (CPT1AD) [MIM:255120]; also known as CPT-I deficiency or CPT1A deficiency. CPT1AD is a rare autosomal recessive metabolic disorder of long-chain fatty acid oxidation characterized by severe episodes of hypoketotic hypoglycemia usually occurring after fasting or illness. Onset is in infancy or early childhood.

  • 序列相似性

    Belongs to the carnitine/choline acetyltransferase family.

  • 细胞定位

    Mitochondrion outer membrane.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • Carnitine O palmitoyltransferase 1 liver isoform antibody
    • Carnitine O palmitoyltransferase I antibody
    • Carnitine O palmitoyltransferase I liver isoform antibody
    • Carnitine O-palmitoyltransferase 1 antibody
    • Carnitine O-palmitoyltransferase I antibody
    • Carnitine palmitoyltransferase 1A (liver) antibody
    • Carnitine palmitoyltransferase 1A antibody
    • Carnitine palmitoyltransferase I antibody
    • Carnitine palmitoyltransferase I liver antibody
    • CPT 1 antibody
    • CPT I antibody
    • CPT1 antibody
    • CPT1 L antibody
    • CPT1-L antibody
    • Cpt1a antibody
    • CPT1A_HUMAN antibody
    • CPTI antibody
    • CPTI-L antibody
    • L CPT1 antibody
    • liver isoform antibody
    see all

图片

  • Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)
    Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: CPT1A knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: MCF-7 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab128568 observed at 88 kDa. Red - loading control, ab181602, observed at 37 kDa.


    ab128568 specifically detected the expected band for CPT1A in wild-type HAP1 cells. No band was observed in CPT1A knockout cells. Wild-type and CPT1A knockout samples were subjected to SDS-PAGE. ab128568 and ab181602 (loading control to GAPDH) were diluted at 1 μg/mL and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [8F6AE9] (ab128568)
    Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [8F6AE9] (ab128568)
    Immunofluorescent staining of CPT1A in H9C2 cells (rat) using ab128568.
    Fixation: 4% paraformaldehyde PBS fixed for 20 minutes
    Permebilization: 0.1% Triton X-100 PBS for 30 minutes at room temperature while rocking
    Blocking: 2x Sigma Block 0.1% Triton X-100 H2O for 2 hours at room temperature while rocking
    Primary antibodies: Anti-CPT1A antibody (ab128568) 0.5 ug/mL 1x Sigma Block with 0.1% Triton X-100 incubated overnight at 4 °C.
    Washing: 3x 1% NGS 10 minutes/wash.
    Secondary antibodies: Alexa 488 GAM 1:1000 diluted in 1% NGS with 0.1% Triton X-100 PBS incubated for 2 hours at room temperature while rocking.
    Washing: 3x 1% NGS 10 minutes/wash.
    DAPI: 20 ng/mL in 1% NGS, 0.1% Triton X-100 PBS.
    Washing: 1x 1% NGS 10 minutes/wash.
  • Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)
    Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)
    All lanes : Anti-CPT1A antibody [8F6AE9] (ab128568) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Partial Human Recombinant CPT1A protein (ab128569) at 0.1 µg
    Lane 3 : Full Length Human Recombinant OTC protein at 0.1 µg
    Lane 4 : HepG2 (Human hepatocellular carcinoma cell line) Whole Cell Lysate at 20 µg
    Lane 5 : H9C2 (Rat cardiomyoblast cell line) Whole Cell Lysate at 20 µg
    Lane 6 : H4IIE ( Rat hepatoma cell line) Whole Cell Lysate at 20 µg
    Lane 7 : RHM (Rat heart mitochondrial homogenate) at 20 µg
    Lane 8 : HLH (Human liver homogenate) at 20 µg
    Lane 9 : RLH (Rat liver homogenate) at 20 µg
    Lane 10 : MLH (Mouse liver homogenate) at 20 µg

    Secondary
    All lanes : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 88 kDa
    Observed band size: 88 kDa


    Exposure time: 1 minute


    Predicted partial recombinant protein band size : 32 kDa
    Observed band size : 32 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [8F6AE9] (ab128568)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [8F6AE9] (ab128568)
    IHC image of CPT1A staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab128568, 0.5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • Flow Cytometry - Anti-CPT1A antibody [8F6AE9] (ab128568)
    Flow Cytometry - Anti-CPT1A antibody [8F6AE9] (ab128568)
    Overlay histogram showing HeLa cells stained with ab128568 (red line) or no primary antibody (black line). The cells were fixed with 4% paraformaldehyde (15 min) and then permeabilized with 0.1% Triton X-100 in PBS, 3% BSA for 10 min. The cells were then incubated in 3% BSA in PBS for 10 minutes to block non-specific protein-protein interactions followed by the antibody (ab128568, 1µg/mL) for 60 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96871) at 1/1000 dilution for 30 min.
  • Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)
    Western blot - Anti-CPT1A antibody [8F6AE9] (ab128568)
    All lanes : Anti-CPT1A antibody [8F6AE9] (ab128568) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : HHH (Human heart homogenate) RIPA Extract Immunoprecipitated with no primary antibody
    Lane 3 : HHH (Human heart homogenate) RIPA Extract Immunoprecipitated with Anti-CPT1A antibody 8F6AE9 (ab128568)
    Lane 4 : HHH (Human heart homogenate) RIPA Extract 20 µg

    Secondary
    All lanes : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1:10000

    Developed using the ECL technique.

    Performed under non-reducing conditions.

    Predicted band size: 88 kDa


    Exposure time: 1 minute
  • In-Cell ELISA - Anti-CPT1A antibody [8F6AE9] (ab128568)
    In-Cell ELISA - Anti-CPT1A antibody [8F6AE9] (ab128568)
    In-Cell ELISA for Anti-CPT1A antibody (ab128568) stained HeLa cells (human)
    Seeding: HeLa cells seeded in a 1:2 dilution series starting at 60,000 cells/well across Row A, 30,000 cells/well across Row B, etc. Row H contains no cells.
    Table: Example In-Cell ELISA Average Data from shown plate.
    Fixation: 4% paraformaldehyde PBS fixed for 15 minutes
    Permeabilization: 0.3% Triton X-100 PBS for 30 minutes at room temperature while shaking
    Blocking: 2x Sigma Block 0.3% Triton X-100 H2O for 2 hours at room temperature while shaking
    Primary antibodies: All primaries diluted in 1x Sigma Block with 0.3% Triton X-100 incubated overnight at 4 °C.
    • Columns 1-3: Anti-CPT1A antibody (ab128568) 10 ug/mL
    • Columns 4-6: Anti-CPT1A antibody (ab128568) 1 ug/mL
    • Columns 7-9: Anti-CPT1A antibody (ab128568) 0.1 ug/mL
    • Columns 10-12: No Primary
    Washing: Briefly 4x with 0.3% TWEEN-20 PBS

文献 (179)

发表研究结果有使用 ab128568？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab128568 被引用在 179 文献中.

  • Rossi T  et al. BETi enhance ATGL expression and its lipase activity to exert their antitumoral effects in triple-negative breast cancer (TNBC) cells. J Exp Clin Cancer Res 42:7 (2023). PubMed: 36604676
  • Guo Z  et al. Psoraleae Fructus Ethanol Extract Induced Hepatotoxicity via Impaired Lipid Metabolism Caused by Disruption of Fatty Acid β-Oxidation. Oxid Med Cell Longev 2023:4202861 (2023). PubMed: 36647431
  • Hai Y  et al. Icariin Alleviates Nonalcoholic Fatty Liver Disease in Polycystic Ovary Syndrome by Improving Liver Fatty Acid Oxidation and Inhibiting Lipid Accumulation. Molecules 28:N/A (2023). PubMed: 36677577
  • Talari NK  et al. Lipid-droplet associated mitochondria promote fatty-acid oxidation through a distinct bioenergetic pattern in male Wistar rats. Nat Commun 14:766 (2023). PubMed: 36765117
  • Yau WW  et al. Nicotinamide riboside rescues dysregulated glycolysis and fatty acid β-oxidation in a human hepatic cell model of citrin deficiency. Hum Mol Genet 32:1922-1931 (2023). PubMed: 36881658
View all Publications for this product

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1-8 of 8 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CPT1A antibody [8F6AE9]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Appendix)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH9.2
Permeabilization
Yes - Tween-20
Specification
Appendix
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Formaldehyde
Read More
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

提交于 Sep 16 2022

Western blot abreview for Anti-CPT1A antibody [8F6AE9]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Tissue lysate - whole (Placenta)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
50 µg
Specification
Placenta
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More
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. Polina Vishnyakova

Verified customer

提交于 Apr 02 2019

Western blot abreview for Anti-CPT1A antibody [8F6AE9]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
human hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More
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

提交于 Jan 25 2018

Western blot abreview for Anti-CPT1A antibody [8F6AE9]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Horse Cell lysate - other (Liver, mitochondrial extract)
Gel Running Conditions
Reduced Denaturing (12.5 % acrylamide)
Loading amount
60 µg
Specification
Liver, mitochondrial extract
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: 22°C
Read More
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

提交于 Jul 22 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-CPT1A antibody [8F6AE9]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Sample
Mouse Cell (Mouse alveolar cells)
Specification
Mouse alveolar cells
Permeabilization
Yes - 90% methanol
Fixative
Formaldehyde
Read More
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

提交于 Jul 23 2013

Question

I've done many mitochondria isolation in liver and it works. So, I just wanted to know if there is a difference in mitochondria isolation in the heart.
.com

Read More
Answer

Hello. I have contacted the laboratory regarding differences between mitchondria isolation in the heart and they have said there is not many differences. Mitochondria preparations are typically better from the heart (purer and higher yield).Heart contains about 30% mitochondria to begin with. Dissection and homogenization are critical because the muscle is not a soft tissue. Buffers are the same. Please let me know if I can be of further assistance.

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Question

I think my mitochondria isolation is not ok. And I want to know if you have any protocol for that.

Read More
Answer



We do have a mitochondria isolation kit for Tissue (ab110168) that you may be interested in https://www.abcam.com/mitochondria-isolation-kit-for-tissue-ab110168.html

Read More

Question

Thank you for your email.
Then, ab128568 and ab22604 are applicale to liver sample?
Please check one more time.


Best regards,

Read More
Answer

Thank you for your inquiry.

Yes, I can confirm that for both of these antibodies liver samples are a good positive control.

ab22604: This antibody gave a positive signal in Human, Rat and mouse liver tissue lysates.

ab128568: Please use one ofthe following aspositive control:HepG2, H9C2, H4IIE Whole Cell Lysates; Rat heart mitochondrial, Human liver, Rat liver and Mouse liver homogenates;

Please see the datasheets for moreinformation:

https://www.abcam.com/Glutathione-Peroxidase-1-antibody-ab22604.html (or use the following: https://www.abcam.com/Glutathione-Peroxidase-1-antibody-ab22604.html).

https://www.abcam.com/CPT1A-antibody-8F6AE9-ab128568.html (or use the following: https://www.abcam.com/CPT1A-antibody-8F6AE9-ab128568.html).

I wish your customer good luck with their experiments.

Read More

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