Overview

  • Product name
    Anti-Leptin Receptor antibody
    See all Leptin Receptor primary antibodies
  • Description
    Rabbit polyclonal to Leptin Receptor
  • Host species
    Rabbit
  • Specificity
    Will detect all isoforms of the leptin receptor.
  • Tested applications
    Suitable for: IP, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

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

    KEIQWKTHEVFDAKSKSA


    (Peptide available as ab5837)

  • Positive control
    • Mouse lung and brain extract, 293 whole cell lysate, Huvec cell lysate.
  • General notes

    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.

Properties

Applications

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.

Application Abreviews Notes
IP Use at an assay dependent concentration. PubMed: 21151569
IHC-Fr Use at an assay dependent concentration.
WB 1/2000. Detects a band of approximately 100, 125 kDa.Can be blocked with Leptin Receptor peptide (ab5837). Corresponding to the long and short forms (respectively) of leptin receptor OB-R from mouse brain extract. Two smaller bands of approximately 31 and 35 kDa are also detected, which cannot be competed out with immunogenic peptide. Block with 5% milk in TBST at RT. The native leptin receptor is a homodimer held together with disulfide bonds. On a reducing SDS-PAGE, bands may appear as approximately 60 kDa subunits. Further variations in size is possible and is due to glycosylation.

Target

  • Function
    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.
  • Tissue specificity
    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.
  • Sequence similarities
    Belongs to the type I cytokine receptor family. Type 2 subfamily.
    Contains 4 fibronectin type-III domains.
    Contains 1 Ig-like (immunoglobulin-like) domain.
  • Domain
    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.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Secreted and Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • All lanes : Anti-Leptin Receptor antibody (ab5593) at 1/2000 dilution

    Lane 1 : Mouse lung whole cell lysate
    Lane 2 : Mouse brain whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Polyclonal goat anti-rabbit conjugated to HRP at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Additional bands at: 100 kDa (possible isoform)


    Exposure time: 10 seconds


    Blocking agent: 5% Milk for 1 hour at 23°C.

    See Abreview

  • Western blot of both short and long forms of Leptin Receptor on mouse lung tissue extract using ab5593.

References

This product has been referenced in:
  • Yuan F  et al. Leptin Signaling in the Carotid Body Regulates a Hypoxic Ventilatory Response Through Altering TASK Channel Expression. Front Physiol 9:249 (2018). Read more (PubMed: 29636698) »
  • Ye C  et al. Leptin alleviates intestinal mucosal barrier injury and inflammation in obese mice with acute pancreatitis. Int J Obes (Lond) 42:1471-1479 (2018). Read more (PubMed: 29934609) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 27 Abreviews or Q&A

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
Abcam has not validated the combination of species/application used in this Abreview.
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

Kevin Zhang

Verified customer

Submitted May 09 2017

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

Abcam user community

Verified customer

Submitted Aug 11 2015

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

Abcam user community

Verified customer

Submitted Jan 27 2014

Abcam has not validated the combination of species/application used in this Abreview.
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

Dr. Brian Lam

Verified customer

Submitted Feb 21 2013

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

Mrs. Judith Kranz

Verified customer

Submitted Feb 19 2013

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

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

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
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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