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CPT1A Antibody - middle region (ARP44796_P050)

100 ul
$319.00
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Conjugation Options

ARP44796_P050-FITC Conjugated

ARP44796_P050-HRP Conjugated

ARP44796_P050-Biotin Conjugated

Gene Symbol:
CPT1A
Official Gene Full Name:
Carnitine palmitoyltransferase 1A (liver)
NCBI Gene Id:
1374
Protein Name:
Carnitine O-palmitoyltransferase 1, liver isoform
Swissprot Id:
P50416
Protein Accession #:
NP_001027017
Nucleotide Accession #:
NM_001031847
Alias Symbols:
CPT1, CPT1-L, L-CPT1
Replacement Item:
This antibody may replace item sc-20514 from Santa Cruz Biotechnology.
Description of Target:
The mitochondrial oxidation of long-chain fatty acids is initiated by the sequential action of carnitine palmitoyltransferase I (which is located in the outer membrane and is detergent-labile) and carnitine palmitoyltransferase II (which is located in the inner membrane and is detergent-stable), together with a carnitine-acylcarnitine translocase. CPT I is the key enzyme in the carnitine-dependent transport across the mitochondrial inner membrane and its deficiency results in a decreased rate of fatty acid beta-oxidation.The mitochondrial oxidation of long-chain fatty acids is initiated by the sequential action of carnitine palmitoyltransferase I (which is located in the outer membrane and is detergent-labile) and carnitine palmitoyltransferase II (which is located in the inner membrane and is detergent-stable), together with a carnitine-acylcarnitine translocase. CPT I is the key enzyme in the carnitine-dependent transport across the mitochondrial inner membrane and its deficiency results in a decreased rate of fatty acid beta-oxidation. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.
Protein Size (# AA):
756
Molecular Weight:
86kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Affinity Purified
Application:
IHC, WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express CPT1A.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express CPT1A.
Immunogen:
The immunogen is a synthetic peptide directed towards the middle region of human CPT1A
Predicted Species Reactivity:
Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish
Tested Species Reactivity:
Human
Predicted Homology Based on Immunogen Sequence:
Cow: 100%; Dog: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 100%; Rabbit: 100%; Rat: 100%; Sheep: 100%; Zebrafish: 100%
Complete computational species homology data:
Anti-CPT1A (ARP44796_P050)
Peptide Sequence:
Synthetic peptide located within the following region: LSTSQTPQQQVELFDLENNPEYVSSGGGFGPVADDGYGVSYILVGENLIN
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Protein Interactions:
UBC; SUMO2; LGR4; RNF2; FBXO6; PARK2; LYN; CLN3; HDAC11; KBTBD7; NPDC1; HSPA5; NR4A1; CYBA; CLIC1; SNRNP200; ERLIN1; TOMM40; EIF3C; PCBP1; NDUFV1; NDUFS1; NDUFA2; LPP; CPS1; ATP6V1C1; RHOA; PCK1; BCL2;
Blocking Peptide:
For anti-CPT1A (ARP44796_P050) antibody is Catalog # AAP44796 (Previous Catalog # AAPP12272)
Datasheets/Manuals:
Printable datasheet for anti-CPT1A (ARP44796_P050) antibody
Target Reference:
Roomets,E., (2008) Invest. Ophthalmol. Vis. Sci. 49 (4), 1660-1664
Publications:

Convertini, P; Menga, A; Andria, G; Scala, I; Santarsiero, A; Castiglione Morelli, MA; Iacobazzi, V; Infantino, V; The contribution of the citrate pathway to oxidative stress in Down syndrome. 149, 423-431 (2016). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 27502741

Laghezza, A. et al. New 2-(aryloxy)-3-phenylpropanoic acids as peroxisome proliferator-activated receptor -alpha/γ dual agonists able to upregulate mitochondrial carnitine shuttle system gene expression. J. Med. Chem. 56, 60-72 (2013). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 23171045

Piemontese, L; Cerchia, C; Laghezza, A; Ziccardi, P; Sblano, S; Tortorella, P; Iacobazzi, V; Infantino, V; Convertini, P; Dal Piaz, F; Lupo, A; Colantuoni, V; Lavecchia, A; Loiodice, F; New diphenylmethane derivatives as peroxisome proliferator-activated receptor alpha/gamma dual agonists endowed with anti-proliferative effects and mitochondrial activity. 127, 379-397 (2017). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 28076827

Product Reviews

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2 Item(s)

02/01/2017 16:23
  • Overall Experience:
  • Quality:
Product Review: CPT1A antibody-middle region (ARP44796_P050) in Human Pancreatic cancer cell lines HPAF1 and Capan1 using Western Blot
Product Page for CPT1A antibody-middle region (ARP44796_P050)

Researcher: Dr. Pankaj Singh, UNMC, Omaha, NE
Application: Western blotting
Species + Tissue/Cell type: Human Pancreatic cancer cell lines HPAF1 and Capan1
How many ug's of tissue/cell lysate run on the gel:
1: 45ug capan1 cell lysate
2: 45 ug HPAF cell lysate
Primary antibody dilution: 1:1000
Secondary antibody: Anti-Rabbit HRP
Secondary antibody dilution: 1:5000



Questionnaire:
How do Aviva's reagents play a role in your experimental goals?
Used in different biochemical characterization.

How would you rate this antibody on a scale from 1-5 (5=best) and why?
5/5, Giving specific band.

Would you use this antibody in future experiments?
Yes.

Have you used another antibody which has worked in your application?
No.

Do you believe the information about the reagent on Aviva's website is correct?
Almost correct.

If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?
We are planning to use it for some of the future experiments

How did you store the antibody after re-suspension?
At -20C in small aliquots.

Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):
Human, Pancreatic cancer cell lines: HPAF1 and Capan1, 45 ug.

How many different experimental trials were conducted using the antibody sample?
1.

How was this sample prepared?
Cells were lysed in RIPA buffer.

Primary antibody dilution and incubation time:
1:1000, Overnight at 4C.

Secondary antibody used and dilution and incubation time:
1:5000, 2 hour, RT.

What controls were used in your experiment (positive/negative)?
Beta-tubulin.

Please include your detailed WB Procedure/Protocol here:
IMMUNOBLOTTING PROTOCOL
1- Prepared the 10 % SDS-PAGE gel.
2- Loaded 45 ug of total protein (Cell lysate) along with molecular weight markers.
3- Run the gel initially at 70V, till the dye reached up to the separating gel and then at 110V till the time dye reached to bottom.
4- Cut PVDF membrane to the same size of the gel. Soaked the membrane in 100% Methanol for 10sec then in water for 20 sec and finally 20 min in transfer buffer.
5- Arranged the Electro-transfer apparatus with gel-membrane sandwich and connected to the power supply at constant Amp (225 mA) for 1hour.
6- Disconnected the transfer apparatus and transferred the membrane in blocking solution [5% Skimmed in PBST (0.1%)] for 1 hour.
7- After blocking transferred the membrane in Primary antibody solution (1:1000 dilutions in PBST) and incubated at 4C for over-night.
8- Washed the membrane 3 times with PBST (0.1%) for 10 minutes each.
9- Incubated the membrane with respective secondary antibody (1:5000 dilutions in PBST) for 2 hour at room temperature.
10- Washed the membrane 3 times with PBST (0.1%) for 10 minutes each.
11- Developed the blots using standard protocol.

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02/01/2017 14:23
  • Overall Experience:
  • Quality:
Product Review: CPT1A antibody-middle region (ARP44796_P050) in Human Capan1 cells (Pancreatic cancer cell line) using IHC

Product Page for CPT1A antibody-middle region (ARP44796_P050)

Researcher: Dr. Pankaj Singh, UNMC, Omaha, NE
Application: IHC
Species + Tissue/Cell type: Human Capan1 cells (Pancreatic cancer cell line)
How many ug's of tissue/cell lysate run on the gel:
Human Capan1 cells (Pancreatic cancer cell line)                                                                                                                Primary antibody dilution: 1:300                                                                                                                                        Secondary antibody: Anti-rabbit-AlexaFluor-488                                                                                                              Secondary antibody dilution: 1:200

 
 
Questionnaire:
How do Aviva's reagents play a role in your experimental goals?
Used in different biochemical characterization.

How would you rate this antibody on a scale from 1-5 (5=best) and why?
4/5, Giving specific band.

Would you use this antibody in future experiments?
Yes.

Have you used another antibody which has worked in your application?
No.

Do you believe the information about the reagent on Aviva's website is correct?
Most of the time its correct.

If the antibody works, do you plan to use it in future experiments or to publish your data?
We are planning to use it for some of the future experiments.
How did you store the antibody after re-suspension?
At -20C in small aliquots.

Sample Description (please include species type and tissue/cell type):
Pancreatic cancer cell line Capan1

Please explain fixation solution/method used (formalin, periodate-lysine-paraformaldehyde, acetone, etc.)?
With 4% Paraformaldehyde
How many different experimental trials were conducted using the antibody sample?
1

Primary antibody dilution, incubation time and temperature:
1:300, 2 hour, RT.
Secondary antibody used, dilution, incubation time and temperature:
1: 200, 1:30 hour, RT

From your IHC/ICC images, briefly explain the colors of each stain and counterstain:
Green- Alexa flour-488, Blue-DAPI
Did you use an antigen retrieval method? If so, please explain?
What controls were used in your experiment?
Secondary Antibody control.

Please include your detailed tissue preparation and staining procedure/protocol here:
1. Seeded 250,000 cells on coverslips in 12 well plate and grown over night
2. Washed the cells with 1X PBS.
3. Fixed the cell in 4% Paraformaldehyde for 15 minute at RT.
4. Incubated the cells with 0.1M Glycine solution for 15 minute at RT to quench the excess paraformaldehyde.
5. Washed the cells with 1X PBS twice and left them in 1% BSA in PBS for over-night.
6. Incubated the cells in 0.1% Triton-X 100 in PBS for 10 minute to permeabilize the cells.
7. Washed the cells with 1X PBS three times.
8. Incubated the cells with DMEM for 5 minute.
9. Added 1° Antibody with 1:300 dilutions in 1% BSA in PBST (0.1%) and incubated at RT for 2 hour.
10. Washed the cells with 1X PBS three times for 5 minutes each.
11. Added 2° Antibody (Alexa Fluor-488, Jackson Immuno Research Laboratories, Inc) with 1:200 dilution in 1% BSA in PBST(0.1%) and incubated at RT for 1:30 hour.
12. Washed the cells with 1X PBS three times for 5 minutes each.
13. Added the Anti-fade with DAPI on glass slides.
14. Mounted the coverslips (cells facing downward) on the glass slide gently.
15. Absorbed the extra liquid with tissue paper.
16. Sealed the edges of coverslips with the help of nail paint.
17. Stored in the slide box at 4C.
18. Captured the image.

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