製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Leptin
  • アプリケーション
    適用あり: ICC/IF, IP, WB, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Sheep, Rabbit, Human, Pig
    交差が予測される動物種: Goat, Horse, Cow, Cat, Dog, Chimpanzee, Macaque monkey, Gorilla
  • 免疫原

    Synthetic peptide:

    SRNVIQISNDLENLRD

    , corresponding to amino acids 91-106 of Human Leptin.

  • ポジティブ・コントロール
    • Human Serum

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

Anti-Leptin antibody 画像

  • 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.

  • Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 3 minutes
  • 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.

Anti-Leptin antibody (ab16227) 使用論文

This product has been referenced in:
  • 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) »
  • Zhang KY  et al. Clinical evaluation of salivary carbohydrate antigen 125 and leptin in controls and parotid tumours. Oral Dis 22:630-8 (2016). Read more (PubMed: 27195940) »

See all 9 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

投稿 Apr 28 2014

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 u...

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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 pr...

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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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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 pe...

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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
Username

Abcam user community

Verified customer

投稿 Feb 24 2012

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 sug...

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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 ...

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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 pre...

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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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