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    leptin-antibody-ab16227.pdf

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Anti-Leptin antibody (ab16227)

  • Datasheet
  • SDS
Reviews (3)Q&A (7)References (19)

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Immunocytochemistry/ Immunofluorescence - Anti-Leptin antibody (ab16227)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Leptin antibody (ab16227)
  • Western blot - Anti-Leptin antibody (ab16227)
  • Immunocytochemistry/ Immunofluorescence - Anti-Leptin antibody (ab16227)
  • Immunocytochemistry/ Immunofluorescence - Anti-Leptin antibody (ab16227)

Key features and details

  • Rabbit polyclonal to Leptin
  • Suitable for: ICC/IF, WB, IHC-P
  • Reacts with: Mouse, Human, Recombinant fragment
  • Isotype: IgG

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Overview

  • Product name

    Anti-Leptin antibody
    See all Leptin primary antibodies
  • Description

    Rabbit polyclonal to Leptin
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Mouse
    Human
    IHC-P
    Human
    WB
    Recombinant fragment
    See all applications and species data
  • Immunogen

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

    SRNVIQISNDLENLRD


    Database link: P41159
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • Human Serum

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Ammonium Sulphate Precipitation
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Growth Factors/Hormones
    • Hormones
    • Neuroscience
    • Neurology process
    • Metabolism
    • Stem Cells
    • Mesenchymal Stem Cells
    • Adipogenesis
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Metabolism
    • Types of disease
    • Obesity
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

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.

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.

Application Species
ICC/IF
Mouse
Human
IHC-P
Human
WB
Recombinant fragment
All applications
Goat
Horse
Cow
Cat
Dog
Chimpanzee
Macaque monkey
Gorilla
Application Abreviews Notes
ICC/IF
1/20 - 1/200.
WB (1)
1/500 - 1/5000. Detects a band of approximately 16 kDa.
IHC-P (1)
Use at an assay dependent concentration.
Notes
ICC/IF
1/20 - 1/200.
WB
1/500 - 1/5000. Detects a band of approximately 16 kDa.
IHC-P
Use at an assay dependent concentration.

Target

  • Function

    May function as part of a signaling pathway that acts to regulate the size of the body fat depot. An increase in the level of LEP may act directly or indirectly on the CNS to inhibit food intake and/or regulate energy expenditure as part of a homeostatic mechanism to maintain constancy of the adipose mass.
  • Involvement in disease

    Defects in LEP may be a cause of obesity (OBESITY) [MIM:601665]. It is a condition characterized by an increase of body weight beyond the limitation of skeletal and physical requirements, as the result of excessive accumulation of body fat.
  • Sequence similarities

    Belongs to the leptin family.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P41159 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 493838 Cat
    • Entrez Gene: 449638 Chimpanzee
    • Entrez Gene: 280836 Cow
    • Entrez Gene: 3952 Human
    • Entrez Gene: 16846 Mouse
    • Omim: 164160 Human
    • SwissProt: Q9N2C1 Cat
    • SwissProt: O02750 Chimpanzee
    • SwissProt: P50595 Cow
    • SwissProt: Q257X2 Goat
    • SwissProt: Q95189 Gorilla
    • SwissProt: Q9TU09 Horse
    • SwissProt: P41159 Human
    • SwissProt: P41160 Mouse
    • Unigene: 194236 Human
    • Unigene: 277072 Mouse
    see all
  • Alternative names

    • FLJ94114 antibody
    • LEP antibody
    • LEP_HUMAN antibody
    • LEPD antibody
    • Leptin (murine obesity homolog) antibody
    • Leptin (obesity homolog, mouse) antibody
    • Leptin antibody
    • Leptin Murine Obesity Homolog antibody
    • Leptin Precursor Obesity Factor antibody
    • OB antibody
    • Obese protein antibody
    • Obese, mouse, homolog of antibody
    • Obesity antibody
    • Obesity factor antibody
    • Obesity homolog mouse antibody
    • Obesity Murine Homolog Leptin antibody
    • OBS antibody
    • OTTHUMP00000212285 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Leptin antibody (ab16227)
    Immunocytochemistry/ Immunofluorescence - Anti-Leptin antibody (ab16227)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Leptin antibody (ab16227)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Leptin antibody (ab16227)
    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.
  • Western blot - Anti-Leptin antibody (ab16227)
    Western blot - Anti-Leptin antibody (ab16227)
  • Immunocytochemistry/ Immunofluorescence - Anti-Leptin antibody (ab16227)
    Immunocytochemistry/ Immunofluorescence - Anti-Leptin antibody (ab16227)

    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/ Immunofluorescence - Anti-Leptin antibody (ab16227)
    Immunocytochemistry/ Immunofluorescence - Anti-Leptin antibody (ab16227)

    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.

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (19)

Publishing research using ab16227? Please let us know so that we can cite the reference in this datasheet.

ab16227 has been referenced in 19 publications.

  • Zhang J  et al. Dlgap1 negatively regulates browning of white fat cells through effects on cell proliferation and apoptosis. Lipids Health Dis 19:39 (2020). PubMed: 32169116
  • Nwadozi E  et al. Leptin is a physiological regulator of skeletal muscle angiogenesis and is locally produced by PDGFRa and PDGFRß expressing perivascular cells. Angiogenesis 22:103-115 (2019). PubMed: 30121753
  • Kong Y  et al. The Effect of the Leptin and Leptin Receptor Expression on the Efficacy of Neoadjuvant Chemotherapy in Breast Cancer. Med Sci Monit 25:3005-3013 (2019). PubMed: 31015393
  • Lo KA  et al. Adipocyte Long-Noncoding RNA Transcriptome Analysis of Obese Mice Identified Lnc-Leptin, Which Regulates Leptin. Diabetes 67:1045-1056 (2018). PubMed: 29519872
  • 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). PubMed: 29436483
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 10 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-Leptin antibody

Poor
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Dec 21 2017

Western blot abreview for Anti-Leptin antibody

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Apr 28 2014

Question

Thank you very much for your reply.

I understand that the immunohystochemistry image about leptin protein localization in the human placenta comes from an end-user, however, as I found it on your website I assume that you agree with the user's results. Therefore my question about where we can expect the staining for leptin, in the cytoplasm or in the nucleus. I am wondering what your characterisation lab says about the leptin antibody and its cellular localization in immunohystochemistry. I would appreciate if you could let me know their opinion.

Also, thank you for letting me know about the AbTrial program. I would be glad to try this antibody on my canine samples and submit the Abreview. However, on the website it stands that you ship at a concentration of 50microL. Is the amount indeed only 50 microL? What is the IgG concentration in it?

Would it be possible to receive a complimentary aliquote sample from you in order to test it in our laboratory on canine reproductive tissues? I would be happy to write the Abreview afterwards, and given that our results are satisfactory, buy the antibody to use it in my experiment. It is planned to have at least one publication from this experiment, and this could be the first one using your antibody in canine reproductive tissues.

Thank you for your consideration and your reply in advance.

Sincerely,

Read More

Abcam community

Verified customer

Asked on Oct 31 2012

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

Abcam Scientific Support

Answered on Oct 31 2012

Question

I am planning on an experiment with canine tissues to detect leptin by immunohystochemistry in paraffin embedded sections and by Western blot.
I have looked at your product "Anti-Leptin antibody (ab16227)" that is predicted to work with dog samples. I was however surprised to see on the images of the website that leptin was shown within the human placenta in the nuclei (!) of the cells. That is very particular, because in the literature leptin has only been detected in the cytoplasma of the cells of different species and of all kinds of tissues.

How do you explain your findings of nucleus staining with your antibody? Which tissue you used as a positive control and were there staining in the nuclei as well?

I would be very glad to get an answer from you, because I would be happy to buy your product, but I would like to know if it does work and whether it does detect the leptin protein.

Thank you very much.

Read More

Abcam community

Verified customer

Asked on Oct 24 2012

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

Abcam Scientific Support

Answered on Oct 24 2012

Question

Dear All,
a customer of our is building a study to compare healty and obese patients.
She is interested in using your ab3583 and ab16227 on human chondrocyte.
Do you have any experitise with those Abs on this specific samples? In particular the customer would like to know if all the leptin is secreted or she can detect the leptin also in cellular lysate (she remove the cell supernatant) with your Abs?

Thanks in advance
and kind regards

Read More

Abcam community

Verified customer

Asked on Jul 09 2012

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

Abcam Scientific Support

Answered on Jul 09 2012

Question


Thank you for the response, well im not really an expect in doing
ELISAs but i do have a little bit of a background however i will need
your help to help me choose the right antibody.
I think the one you suggested will work best
(https://www.abcam.com/Leptin-antibody-ab16227.html) but tell me does
it give color change at the end?

Read More

Abcam community

Verified customer

Asked on Jun 08 2012

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 Scientific Support

Answered on Jun 08 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Leptin antibody

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Feb 24 2012

Question

Dear technical team,   Our customer would like to purchase ab62551 and ab16227. She is going to test IHC-P with human sample and needs positive control for those two antibodies. How about ab4616 for ab62551 and ab4360 for ab16227?   Please check this for our customer.   Thank you.   Best regards,

Read More

Abcam community

Verified customer

Asked on Jul 29 2011

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

Abcam Scientific Support

Answered on Jul 29 2011

Question

can i use human leptin elisa kit for cattle leptin?

Read More

Abcam community

Verified customer

Asked on Dec 18 2005

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

Abcam Scientific Support

Answered on Dec 19 2005

Question

I am Dr. ?ak?ro?lu from Turkey. I would like to know if you have cow leptin antibody suitable to detect cow plasma leptin using ELISA.The product code I mentioned above, ? guess is not suitable for ELISA, but I just wrote in order to be able to send the mail. I will be very glad with your information. Thanks for sparing your time

Read More

Abcam community

Verified customer

Asked on Nov 28 2005

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.

Read More

Abcam Scientific Support

Answered on Nov 28 2005

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