Overview

  • Product name

    Anti-Lrp2 / Megalin antibody
    See all Lrp2 / Megalin primary antibodies
  • Description

    Rabbit polyclonal to Lrp2 / Megalin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ELISA, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptides derived from the C terminal part of Human Lrp2/ Megalin.

Applications

Our Abpromise guarantee covers the use of ab101011 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
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    Acts together with cubilin to mediate HDL endocytosis (By similarity). May participate in regulation of parathyroid-hormone and para-thyroid-hormone-related protein release.
  • Tissue specificity

    Absorptive epithelia, including renal proximal tubules.
  • Involvement in disease

    Defects in LRP2 are the cause of Donnai-Barrow syndrome (DBS) [MIM:222448]; also known as faciooculoacousticorenal syndrome (FOAR syndrome). DBS is a rare autosomal recessive disorder characterized by major malformations including agenesis of the corpus callosum, congenital diaphragmatic hernia, facial dysmorphology, ocular anomalies, sensorineural hearing loss and developmental delay. The FOAR syndrome was first described as comprising facial anomalies, ocular anomalies, sensorineural hearing loss, and proteinuria. DBS and FOAR were first described as distinct disorders but the classic distinguishing features between the 2 disorders were presence of proteinuria and absence of diaphragmatic hernia and corpus callosum anomalies in FOAR. Early reports noted that the 2 disorders shared many phenotypic features and may be identical. Although there is variability in the expression of some features (e.g. agenesis of the corpus callosum and proteinuria), DBS and FOAR are now considered to represent the same entity.
  • Sequence similarities

    Belongs to the LDLR family.
    Contains 17 EGF-like domains.
    Contains 36 LDL-receptor class A domains.
    Contains 37 LDL-receptor class B repeats.
  • Cellular localization

    Membrane. Membrane > coated pit.
  • Information by UniProt
  • Database links

  • Alternative names

    • Calcium sensor protein antibody
    • DBS antibody
    • Glycoprotein 330 antibody
    • gp330 antibody
    • Heymann nephritis antigen homolog antibody
    • Low-density lipoprotein receptor-related protein 2 antibody
    • LRP-2 antibody
    • Lrp2 antibody
    • LRP2_HUMAN antibody
    • Megalin antibody
    see all

References

This product has been referenced in:

  • Baines RJ  et al. CD36 mediates proximal tubular binding and uptake of albumin and is upregulated in proteinuric nephropathies. Am J Physiol Renal Physiol 303:F1006-14 (2012). Read more (PubMed: 22791331) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunoprecipitation
Sample
Rat Cell lysate - whole cell (Primary Neurons)
Total protein in input
250 µg
Specification
Primary Neurons
Immuno-precipitation step
Other - Dynabeads Protein A

Abcam user community

Verified customer

Submitted Jul 13 2012

Answer

Thank you for providing that extra information.

The band that can be expected with this lysate is the predicted 517 kDa, however, to our knowledge, this lysate has not been specifically probed for Lrp2 and I cannot therefore be sure of this. Additionally, ab56014 has not been used to detect the endogenously expressed protein in western blotting in house (which is also seems to betrue of the Sc-25470). Only recombinantly expressed protein fragments have been detected. This makes performing the western blot considerably easier as the transfer of the smaller recombinant protein is much less problematic than transferring a 517 kDa protein.

Can I ask, are you seeing any bands after performing the western blot or is the membrane completely blank? If you could possibly share an image of the blot obtained that would be very helpful. What was the result of the "no primary" control?

I would advise tryingthe following in order to improve the resultsobserved to date:

1.I would prepare the protein samples by heating for 10 minutes to 70 degrees. If you are not already doing so I would also used a reducing agent in the loading buffer. This is not necessary with the control lysate as this is already included (butthe heating step should still be performed).

2. In order to improve the transfer and binding of the protein to the membrane I would use PVDF membrane and make sure to pretreat in methanol for 1-2 minutes, followed by incubation in ice cold tranfer buffer for 5 minutes.

3. I would use a very low percentage gel in order to improve the transfer, as well as adding 0.1% SDS to the tranfer buffer as this prevents the precipitation of the protein in the gel. I would also remove the methanol from the transfer buffer as this tends to remove the SDS. Methanol is only necessary in the transfer buffer if using a nitrocellulose membrane. Removing the methanol also promotes the swelling of the membrane which allows large proteins to transfer more easily.

I hope these suggestions improve the results observed so far. Performing a western blot with such a large protein will be quite difficult and may require quite a lot of optimisation in order to get a good result.

I look forward to receiving yourreply.

Read More

Answer

Thank you for your enquiry regarding ab101011.

The peptides are selected from the cytoplasmic domain: topological domain: 4447 – 4655.

I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More

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