Anti-Leptin Receptor 抗体 (ab5593)

Reviews (6)Q&A (21)References (21)

Key features and details

  • Rabbit polyclonal to Leptin Receptor
  • Suitable for: WB
  • Reacts with: Mouse
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-Leptin Receptor antibody
    Leptin Receptor 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Leptin Receptor

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WBmore details

  • 種交差性

    交差種: Mouse
    交差が予測される動物種: Rat, Human

  • 免疫原

    Synthetic peptide corresponding to Rat Leptin Receptor aa 577-594 (extracellular).
    Sequence:

    KEIQWKTHEVFDAKSKSA


    (Peptide available as ab5837)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • WB: Mouse lung tissue extract.

  • 特記事項

    As per Uniprot database (P48357) OB receptor has 5 isoforms and is highly glycosylated, this means the observed band size will be different than predicted and multiple bands are expected depending upon the isoforms expressed in sample types.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab5593 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
WB 1/2000. Detects a band of approximately 100, 125 kDa.Can be blocked with Leptin Receptor peptide (ab5837).

ab5593 detects an ~100 kDa and ~125 kDa protein representing the short and long forms of OB-R.

 

ターゲット情報

  • 機能

    Receptor for obesity factor (leptin). On ligand binding, mediates signaling through JAK2/STAT3. Involved in the regulation of fat metabolism and, in a hematopoietic pathway, required for normal lymphopoiesis. May play a role in reproduction. Can also mediate the ERK/FOS signaling pathway.

  • 組織特異性

    Isoform A is expressed in fetal liver and in hematopoietic tissues and choroid plexus. In adults highest expression in heart, liver, small intestine, prostate and ovary. Low level in lung and kidney. Isoform B is highly expressed in hypothalamus.

  • 配列類似性

    Belongs to the type I cytokine receptor family. Type 2 subfamily.
    Contains 4 fibronectin type-III domains.
    Contains 1 Ig-like (immunoglobulin-like) domain.

  • ドメイン

    The cytoplasmic domain may be essential for intracellular signal transduction by activation of JAK tyrosine kinase and STATs.
    The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
    The box 1 motif is required for JAK interaction and/or activation.

  • 翻訳後修飾

    On ligand binding, phosphorylated on two conserved C-terminal tyrosine residues (isoform B only) by JAK2. Tyr-986 is required for complete binding and activation of PTPN11, ERK/FOS activation and, for interaction with SOCS3 (By similarity). Phosphorylation on Tyr-1141 is required for STAT3 binding/activation.

  • 細胞内局在

    Secreted and Cell membrane.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • CD 295 antibody
    • CD295 antibody
    • CD295 antigen antibody
    • Db antibody
    • Fa antibody
    • HuB219 antibody
    • LEP R antibody
    • LEP-R antibody
    • LEPR antibody
    • LEPR_HUMAN antibody
    • LEPRD antibody
    • Leptin receptor antibody
    • Leptin receptor fatty antibody
    • Leptin receptor gene related protein antibody
    • Leptin receptor precursor antibody
    • Leptin receptor precursor antibody
    • OB R gene related protein antibody
    • OB receptor antibody
    • OB-R antibody
    • OB-RGRP antibody
    • obl antibody
    • Obr antibody
    see all

画像

  • Western blot - Anti-Leptin Receptor antibody (ab5593)
    Western blot - Anti-Leptin Receptor antibody (ab5593)
    Anti-Leptin Receptor antibody (ab5593) at 1/2000 dilution

    Western blot detction of both short and long forms of Leptin (OB-R).

参考文献 (21)

ab5593 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab5593 は 21 報の論文で使用されています。

  • van de Looij Y  et al. Nutritional Intervention for Developmental Brain Damage: Effects of Lactoferrin Supplementation in Hypocaloric Induced Intrauterine Growth Restriction Rat Pups. Front Endocrinol (Lausanne) 10:46 (2019). PubMed: 30800096
  • Santamarina AB  et al. Obesity-related inflammatory modulation by juçara berry (Euterpe edulis Mart.) supplementation in Brazilian adults: a double-blind randomized controlled trial. Eur J Nutr N/A:N/A (2019). PubMed: 31197507
  • Fischer J  et al. Leptin signaling impairs macrophage defenses against Salmonella Typhimurium. Proc Natl Acad Sci U S A 116:16551-16560 (2019). PubMed: 31350351
  • Tang XM  et al. Upregulation of suppressor of cytokine signaling 3 ameliorates spinal degenerative disease in adolescents by mediating leptin and tumor necrosis factor-a levels. Exp Ther Med 18:2231-2237 (2019). PubMed: 31410173
  • Chu G  et al. Leptin Receptor Mediates Bmal1 Regulation of Estrogen Synthesis in Granulosa Cells. Animals (Basel) 9:N/A (2019). PubMed: 31683864
View all Publications for this product

レビューと Q&A

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レビューを送る 質問を送る

1-10 of 27 Abreviews or Q&A

Question

I tried to detect the long form of leptin receptor with the newantibody you send to me. But I can detect short form(100KDa) only, but not the long form(125KDa). Would you please find out what is the possible problem for me?


I load about 40μg protein for lung tissue per lane in 7.5% SDS gel. Should I load more? How strong the125KDa band should be compare to the 100KDa band usually?

Electrophoresis: 120V about 2 hours.

Transfer: 400mA for 3hours.

Blocking: 5% dry milk for 1hour.

Incubate with primary antibody(ab5593 1:2000) over night at 4℃.

Rinse with TBST 10 mins ×3times.

Incubate with anti rabbit antibody for one hour at RT.

Rinse with TBST 10 mins ×3times.

ECL

Lung tissues from 2 wide type C57BL6 mice with not treatment and normal chow diet.

My lysis buffer for tissue lysate

50mM HPES(PH 7.6), 150mM NaCl, 20mM Na pyrophoshate, 20mM Beta-Glycerophoshate, 10mM MNaF, 2mM EDTA(PH 8.0), 1% IGEPAL, 10% glycerol, 1mM MgCl2, 1mM CaCl2, 2mM Na orthovanadate, 2mM PMSF, 10μg/ml leupeptin, 10μg/ml aprotinin.

Thank for your help

Read More
Answer

Thank you very much for keeping me updated about the situation, and I'm sorry to hear that the new lot is also not giving the expected results.

As mentioned before, your protocol looks very similar to the protocol used by the lab to test the antibody (attached).In our testing, the 125 kDa band looks fainter than the 100 kDa band, butthe Western blot images in the literature do not show the 100 kDa band so I can not comment on their results.I've reviewed all of the information that I could find on this antibody, and it seemsthat the antibody has been tested with the membrane fractions of the lung and brain lysates (including in our lab testing), so I would suggest trying a membrne fractionation and seeing if this enriches the samples enough to visualize the 125 kDa band. We have a subcellular fraction protocol that could be used-

https://www.abcam.com/index.html?pageconfig=resource&rid=11473

As this protein is heavily glycosylated and also at a higher molecular weight (meaning difficult to transfer), I'd also suggest running a Ponceau stain on the membraneto make sure that the long isoform efficiently transferred.

I'm also wondering if the tissue lysates are whole brain and lung, or are they specific regions of these organs? I can check with the lab whether their samples were taken from the same region.

I look forward to hearing from you. I am very sorry for all of the trouble with this antibody, but I hope to resolve this as quickly as possible. Please let me know if there is anything else that we can do for you.

Read More

Immunohistochemistry (Frozen sections) abreview for Anti-Leptin Receptor antibody

 Poor
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Cell (Hypothalamus)
Permeabilization
Yes - 0.5% Triton X-100
Specification
Hypothalamus
Blocking step
Serum as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Paraformaldehyde
Read More

Kevin Zhang

Verified customer

投稿 May 09 2017

Western blot abreview for Anti-Leptin Receptor antibody

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (mouse lung, brain)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
mouse lung, brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

投稿 Aug 11 2015

Western blot abreview for Anti-Leptin Receptor antibody

 Average
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Denaturing (10)
Sample
Mouse Cell lysate - other (C2C12)
Specification
C2C12
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Read More

Abcam user community

Verified customer

投稿 Jan 27 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Leptin Receptor antibody

 Poor
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary Hypothalamic Neurons)
Specification
Primary Hypothalamic Neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% NP40 in PBS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Dr. Brian Lam

Verified customer

投稿 Feb 21 2013

Western blot abreview for Anti-Leptin Receptor antibody

 Poor
Abreviews
Application
Western blot
Sample
Mouse Tissue lysate - whole (hypothalamus)
Loading amount
25 µg
Specification
hypothalamus
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Mrs. Judith Kranz

Verified customer

投稿 Feb 19 2013

Question

Q. How satisfied are you with how the complaint was handled?
A.Neither satisfied nor unsatisfied

Q. Would you mind if our scientific support team contacted you to better understand what happened and reach an agreeable solution?
A.No, I would not mind

Read More
Answer

Thank you very much for participating in our recent survey.

I am very sorry that the long form of the OBR protein is still not being picked up with this antibody. Have you tried the membrane fractionation to enrich your samples, or the Ponceau stain to verify protein transfer (mentioned in my previous email)? Are you using whole lung and brain lysates, or lysates from specific regions?

We do guarantee these results and I can send a replacement antibody or issue a credit or refund for your original purchase if you would like. Please keep me updated with any new results and let me know if there is anything else that we can do for you.

Read More

Question

My supervisor advised me to ask you send us a free of charge replacement vial from a different lot.


Thank for your help!

Read More
Answer

Thank you very much for your reply.

I'm sending a free of charge vial from a new lot on the order ***, which should arrive tomorrow.

Please keep me updated about the results with this new vial, and if there is anything else that we can do for you please let me know. I'll be happy to help.

Have a nice day!

Read More

Question

Thank for your reply and two kind options. I have to discuss it with my supervisor. My supervisor ison overseas tripnow, once we make the decision,I will let you konw.

Best

Read More
Answer

Thank you very much for your reply, and I apologize for the trouble with this antibody.

I look forward to hearing from you once you have discussed the options with your supervisor. Please let me know if there is anything else that we can do for you.

Have a nice day.

Read More

Question

I'm still waiting for your reply.

Read More
Answer

Thank you very much for your emails and for your patience.

I have been discussing the situation with my colleagues in the lab, and I've found out that the mouse lung lysate used for the WB on our datasheet also came from a healthy, normal-fedmouse.According to the literature (as you have also found), the long form of this receptor is not expressed in obese or diabetic mice, but is expressed in wild type C57Bl/6J mice, so we are surprised that the receptor is not detected in these samples. One of my colleagues has found in the literature, though we have not tested, that mRNA of the long form of the receptor increases during fasting, so this may be one way to boost expression and detection of the 125kD product.Additionally, we loaded 40ug of lysate per lane for detection of both bands in the WB image on the website, which you have also loaded as well. The rest ofour protocol is also very similar to yours, so we are not sure exactly why the long form is not being picked up in your samples.

We do fully guarantee this antibody to work as shown on the datasheet, so I would be happy to send you a free of charge replacement vial from a different lot in order to compare the results. Would you be interested in this option? Alternatively we can issue a credit or refund for your purchase.

I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

Read More

1-10 of 27 Abreviews or Q&A

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