Recombinant
RabMAb

Anti-p75 NGF Receptor antibody [EP1039Y] (ab52987)

Overview

  • Product name
    Anti-p75 NGF Receptor antibody [EP1039Y]
    See all p75 NGF Receptor primary antibodies
  • Description
    Rabbit monoclonal [EP1039Y] to p75 NGF Receptor
  • Host species
    Rabbit
  • Specificity
    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
  • Tested applications
    Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human p75 NGF Receptor aa 350-450. The exact sequence is proprietary.
    Database link: P08138

  • Positive control
    • PC12 cell membrane lysate, PC-12 cell lysate Human brain gilioma tissue
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52987 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
WB 1/50000. Detects a band of approximately 75 kDa (predicted molecular weight: 45 kDa).
IP 1/50.
IHC-P Use at an assay dependent concentration.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. 

ICC/IF 1/50.
Flow Cyt 1/60.

Target

  • Function
    Low affinity receptor which can bind to NGF, BDNF, NT-3, and NT-4. Can mediate cell survival as well as cell death of neural cells.
  • Sequence similarities
    Contains 1 death domain.
    Contains 4 TNFR-Cys repeats.
  • Domain
    Death domain is responsible for interaction with RANBP9.
    The extracellular domain is responsible for interaction with NTRK1.
  • Post-translational
    modifications
    N- and O-glycosylated.
    O-linked glycans consist of Gal(1-3)GalNAc core elongated by 1 or 2 NeuNAc.
    Phosphorylated on serine residues.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD271 antibody
    • CD271 antigen antibody
    • Gp80 LNGFR antibody
    • Gp80-LNGFR antibody
    • Low affinity nerve growth factor receptor antibody
    • Low affinity neurotrophin receptor p75NTR antibody
    • Low-affinity nerve growth factor receptor antibody
    • Nerve growth factor receptor antibody
    • Nerve growth factor receptor TNFR superfamily member 16 antibody
    • NGF receptor antibody
    • Ngfr antibody
    • p75 ICD antibody
    • p75 Neurotrophin receptor antibody
    • p75 NTR antibody
    • p75(NTR) antibody
    • p75NTR antibody
    • TNFR Superfamily Member 16 antibody
    • TNFRSF16 antibody
    • TNR16_HUMAN antibody
    • Tumor necrosis factor receptor superfamily member 16 antibody
    see all

Images

  • Purified ab52987 staining p75 NGF receptor in PC-12 (rat adrenal gland pheochromocytoma) by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 3.9 µg/ml. An AlexaFluor®488 Goat anti-Rabbit was used as the secondary antibody at 2 µg/ml. DAPI was used as a nuclear counterstain. Confocal image showing cytoplasmic and Membranous staining in PC-12 cells.

  • Purified ab52987 staining p75 NGF receptor in paraffin embedded Human tonsil tissue sections by Immunohistochemistry. Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at 3.3μg/ml. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on germinal centre of human tonsil.

  • All lanes : Anti-p75 NGF Receptor antibody [EP1039Y] (ab52987) at 1/1000 dilution (purified)

    Lane 1 : Capan-1 (Human pancreas adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse uterus tissue lysates
    Lane 3 : Mouse brain tissue lysates
    Lane 4 : Rat uterus tissue lysates
    Lane 5 : Rat brain tissue lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 45 kDa


    Exposure time: 180 seconds


    Blocking and diluting buffer: 5% NFDM/TBST

    ab52987 fails to detect band of interest in Capan-1 (positive, PMID: 14613990) and brain lysates (positive, PMID: 21413144, 21541365), indicating its low affinity in some p75 NGF Receptor positive materials.

  • Lane 1 (input): PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 10μg
    Lane 2 (+): PC-12 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52987 in PC-12 whole cell lysate

    Ab52987 immunoprecipitating p75 NGF receptor in PC-12 whole cell lysates. For western blotting, primary antibody used was ab52987 at 1.6 μg/ml. Ab131366 VeriBlot for IP (HRP) was used as the secondary antibody at 1:1000 dilution. Capture antibody was used at 1:40 dilution (2μg in 0.35mg lysates).

    Blocking and diluting buffer: 5% NFDM/TBST.

    Exposure: 10 seconds

  • Flow cytometry analysis of PC-12 (rat adrenal gland pheochromocytoma) cells labeling p75 NGF Receptor with purified ab52987 at 1/80 dilution (1ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • All lanes : Anti-p75 NGF Receptor antibody [EP1039Y] (ab52987) at 1/1000 dilution (purified)

    Lane 1 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Human hippocampus tissue lysates
    Lane 3 : Human brain cortex tissue lysates
    Lane 4 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
    Lane 5 : Mouse hippocampus tissue lysates
    Lane 6 : Mouse cerebral cortex lysates
    Lane 7 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 8 : Rat hippocampus tissue lysates
    Lane 9 : Rat brain cortex tissue lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 45 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

    ab52987 fails to detect band of interest in hippocampus and cortex lysates (positive, PMID: 25180603, 28507518, 18930453, 20937383, 21059364), indicating its low affinity in some p75 NGF Receptor positive materials.

    Exposure: Lane 1-3: 8 seconds
                     Lane 4-6: 180 seconds
                     Lane 7-9: 30 seconds


  • Immunohistochemical analysis of murine uterus tissue with adenomyosis, staining p75 NGF Receptor with ab52987.

    Antigen retrieval was performed by heat mediation in citrate buffer (pH 6). Tissue was blocked with goat serum for 15 minutes before incubating with primary antibody (1/100) overnight at 4°C. A biotinylated goat anti-rabbit IgG was used as the secondary antibody and staining was detected using DAB.
  • ICC/IF image of ab52987 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52987, 1 µg/mL) overnight at 4oC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluo® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.4 µM.

  • Anti-p75 NGF Receptor antibody [EP1039Y] (ab52987) at 1/50000 dilution + PC12 cell lysate at 10 µg

    Secondary
    goat anti-rabbit HRP labelled at 1/2000 dilution

    Predicted band size: 45 kDa
    Observed band size: 75 kDa (why is the actual band size different from the predicted?)

  • Flow Cytometry analysis of PC-12 (rat adrenal gland pheochromocytoma) cells labeling p75 NGF Receptor with unpurified ab52987 at 1/60 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Chang LH  et al. Blockade of soluble epoxide hydrolase attenuates post-ischemic neuronal hyperexcitation and confers resilience against stroke with TrkB activation. Sci Rep 8:118 (2018). Read more (PubMed: 29311641) »
  • Tsai MS  et al. Nerve growth factor upregulates sirtuin 1 expression in cholestasis: a potential therapeutic target. Exp Mol Med 50:e426 (2018). Read more (PubMed: 29328070) »

See all 15 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse neural stem cell)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
40 µg
Specification
mouse neural stem cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Robert Kupp

Verified customer

Submitted Jun 27 2018

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (8)
Sample
Human Cell lysate - whole cell (Melanoma)
Specification
Melanoma
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C
Username

Abcam user community

Verified customer

Submitted May 30 2014

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up