Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab3583 in the following tested applications.

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

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
Inhibition Assay Use at an assay dependent concentration.
ICC 1/50 - 1/200.
IHC-Fr 1/50 - 1/200.
WB 1/200 - 1/1000. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).

Target

Anti-Leptin antibody images

  • Immunohistochemical staining of Leptin in Sheep brain using ab3583.



  • Predicted band size : 16 kDa

    ab3583 at a dilution of 1 / 4000 staining leptin in mouse 3T3-L1 cell lysate by Western blot.

  • ICC/IF image of ab3583 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3583, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Leptin antibody (ab3583)

This product has been referenced in:
  • Johnstone SA  et al. Comparison of human olfactory and skeletal MSCs using osteogenic nanotopography to demonstrate bone-specific bioactivity of the surfaces. Acta Biomater 13:266-76 (2015). Read more (PubMed: 25463488) »
  • Al-Suhaimi E Molecular mechanisms of leptin and pro-apoptotic signals induced by menadione in HepG2 cells. Saudi J Biol Sci 21:582-8 (2014). Read more (PubMed: 25473367) »

See all 6 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Non-Denaturing (Native) (12.5)
Sample Spermophilus tridecemlineatus Tissue lysate - whole (white adipose tissue)
Specification white adipose tissue
Blocking step BSA as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Feb 05 2014

Thank you for contacting us. This antibody has not been tested for blocking functional activity. If you do choose to test it this we way encourage you to submit a review to let us and our customers know whether it worked for you in functional studies....

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Mesenchymal stem cell)
Specification Mesenchymal stem cell
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Username

Mr. Steven Johnstone

Verified customer

Submitted Sep 05 2012

Thank you for your enquiry.

I can confirm that ab3583 and ab16227 antibodiesshould detect leptin both intra and extracellularlly.However based on the references for these product it appears that most if not all of the leptin is immediately s...

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Thank you for passing some information to me.
I understand that ab3583 has been used previously at 1:4000.
Have you had a chance to test this antibody at dilutions of 1/500 or 1/1000 (for 1 hr at room temperature or overnight at 4oC?
Wou...

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Thank you for getting back to me and for specifying the sample types.
As you have kindly explained to me in your previous e-mail, the primary and the secondary antibodies worked with the positive control but the signal was very weak using lysates ...

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Thank you for confirming the order dfetails.
Would you be so kind to clarify what samples you have been using:
- species,
- cell or tissue types,
- lysate (total cell lysates, or cytoplasmic fraction etc)
I look forward to hear...

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Thank you for your prompt answers. Your co-operation in this matter is highly appreciated.
I understand that you have already carried out some optimization steps and it seems that the primary and the secondary antibodies worked on positive control...

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Thank you for enquiry and for taking the time to provide some useful details of the experiments. As I understand three of our products (ab3583, ab13987 and ab97051) have been used in these studies.
Though you have kindly provided some details, it ...

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Thank you for contacting us.

I am sorry that the vial you received did not contain the full amount of the product and I apologize for the inconvenience.We will be pleased to issue a free of charge replacement vial. In order to do this, I am ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"