Overview

  • Product name
  • Description
    Rabbit polyclonal to Leptin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IP, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Sheep, Rabbit, Human, Pig
    Predicted to work with: Goat, Horse, Cow, Cat, Dog, Chimpanzee, Macaque monkey, Gorilla
  • Immunogen

    Synthetic peptide corresponding to Human Leptin aa 91-106.
    Sequence:

    SRNVIQISNDLENLRD


    Database link: P41159

  • Positive control
    • Human Serum

Properties

Applications

Our Abpromise guarantee covers the use of ab16227 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 1/20 - 1/200.
IP Use at an assay dependent concentration.
WB 1/500 - 1/5000. Detects a band of approximately 16 kDa.
IHC-P Use at an assay dependent concentration.

Target

Images

  • Immunocytochemistry/Immunofluorescent analysis of Leptin (green) showing staining in the secretion of NIH-3T3 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab16227 in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 �C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 100x.

  • Ab16227 staining human placenta.
    Left panel: with primary antibody. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • Immunocytochemistry/Immunofluorescent analysis of Leptin (green) showing staining in the secretion of HepG2 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab16227 in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 �C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 100x.

  • Immunocytochemistry/Immunofluorescent analysis of Leptin (green) showing staining in the secretion of HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab16227 in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 �C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 100x.

References

This product has been referenced in:
  • Sun C  et al. Leptin-induced inflammation by activating IL-6 expression contributes to the fibrosis and hypertrophy of ligamentum flavum in lumbar spinal canal stenosis. Biosci Rep 38:N/A (2018). Read more (PubMed: 29436483) »
  • Pan Y  et al. Leptin-LepRb Expressed in Gastric Cancer Patients and Related to Cancer-Related Depression. Biomed Res Int 2017:6482842 (2017). Read more (PubMed: 28316984) »
See all 12 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Mouse stomach)
Permeabilization
Yes - PBS-Tx
Specification
Mouse stomach
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 21 2017

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Spermophilus tridecemlineatus Serum (serum)
Specification
serum
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Apr 28 2014

Answer

Thank you for your reply.

The lab confirmed that leptin is cytoplasmic and not nuclear. The image as well as the legend shown on the product datasheet are therefore incorrect and I asked my colleagues responsible for the website to remove or update the information. Thank you for highlighting this mistake to us.

Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

As explained in my previous email, when a product is to be used in an untested species or application, end users may be eligible for our AbTrial program.

Please reply to this email if you are interested in testing this product and I will send you an AbTrial discount code. After submitting your results the discount code will become active and will be useable on your next order to receive that value off. The Terms and Conditions of this offer can be found at: https://www.abcam.com/abtrial.

Please note that This item is currently out of stock and is estimated to be available by late-November 2012. We apologise for any inconvenience. You may prefer to test another anti-Leptin instead. The link to the list of all Abcam anti-Leptin antibodies is : https://www.abcam.com/index.html?t=58&pt=1.

The last lots of ab3583 we had in stock had a concentration of 0.7 mg/ml, the unit size is indeed 50µl.

I hope this is helpful to you. Do not hesitate to contact me again if you have questions.

Read More

Answer

Thank you very much for your interest in our anti-Leptin ab16227.

The IHC-P image comes from an end-user's feedback, not from the characterisation lab.

We always welcome valuable feedback from our customers, so if you would like to provide your own feedback on this product in IHC-P and with Dog samples, you can participate in our AbTrial program and use our products without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting us know you are interested in testing this product.
2. Our scientists will email you an inactive personal discount code for the value of the product.
3. Purchase and test the product at the regular price.
4. Submit your results, including your discount code in the additional notes section of your Abreview.
5. Once the Abreview is submitted, the discount code will become active.
6. Apply your discount code on your next order to receive that value off.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

Read More

Answer

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 secreted and would therefore be found extracellularlly.

I am sorry that as far as we are aware, these antibodies have not specifically been tested inchondrocyts.However, I would like to reassure you that our antibodies are tested and covered by our 6 month guarantee in the listed tested applications and species on the individual datasheets.

I hope this information is helpful. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

The antibody ab16227 is not conjugated to anything that will produce color. To use this for ELISA, you will need an anti-rabbit IgG antibody, conjugated to an enzyme that will catalyze a color-producing reaction. This enzyme is typically horseradish peroxidase, or HRP for short. One example of such a secondary antibody is ab97064.

However, I am concerned that your meaning for "secondary" is different from mine. Are you trying to do a sandwich ELISA? If so you will need a capture antibody, coated onto the plate, and a detection antibody, which sticks to the leptin that has been captured. If this is what you want to do, the best pair of antibodies for you, assuming your samples are of human origin, will probably be a rabbit polyclonal antibody raised agains the full protein, such as ab13986, and ab6055 for detection, which is directly conjugated to HRP and does not need a secondary anti-IgG.

You will still need a substrate and chromagen for development of color, which we do not have in our catalogue yet.

Please let us know if you have any questions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (small intestine: epithelial cells)
Specification
small intestine: epithelial cells
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate pH=6
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Feb 24 2012

Answer

Thank you for your enquiry. I have just quickly checked the datahseets and the specificity of these products for you and can confirm that these slides can be used as positive control. - ab4616 for ab62551 and - ab4360 for ab16227 I would suggest following the staining procedure (provided in the legend og the IHC image) and perhaps applying signal enhancing step (Dako Envision Flex amplification or ABC system). I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More

Question
Answer

Thank you for your enquiry regarding ab16227 and your interest in our products. This antibody to Leptin has not been tested in Bovine. The sequence between human and bovine leptin only differ by 1 a.a. residue, so I would expect it to work, but we have not tested this product in Bovine. I hope this information helps.

Read More

Answer

Thank you for your enquiry. First, I would like to apologize if I spelled your name incorrectly. It appears that some of the characters in your name did not email properly. It appears that ab16227 is the only leptin antibody we carry that is predicted to react with cow tissue, and as you noted, this antibody has not been tested in ELISA. It is possible that the antibody would work, but we have not tested it. Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will offer you 50 Abpoints which can be redeemed on a number of rewards (a further 100 Abpoints will be offered for an image). I hope this information helps. Please do not hesitate to contact us if you need anything further.

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

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