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

    • ICC/IF: PC-12 cells. IHC-P: Human tonsil tissue; Mouse uterus tissue. WB: Capan-1, SW80, Neuro-2a and PC-12 cell lysate; Mouse uterus, hippocampus and cerebral cortex lysate; Rat uterus, hippocampus and brain cortex lysate; Human hippocampus and brain cortex lysate. IP: PC-12 cell lysate. Flow Cyt: PC-12 cells.
  • 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 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.

  • The differentiation capacity of purified CD34+ cells cultured for 3 days in the presence or absence of ALK5i was evaluated by performing immunofluorescence analysis assessing whether CD34+ cells had changed to cells expressing p75 and/or α-SMA. Expressions of CD34, p75 and α-SMA were assessed by immunofluorescence on day 0 (F-H), day 3 in SP+f medium (I-K), or day 3 in the same medium as (I-K) but with ALK5i (L-N).

    Cultured re-aggregates were fixed in 4% PFA and embedded in paraffin. Sections (5 ∝m) were boiled in 0.01 M citrate (pH 6.0) with 0.1% Tween 20 for 10 min, washed three times in 0.1% Tween-20/PBS, transferred to blocking solution containing 5% BSA and 5% horse serum (Sigma) or goat serum (Invitrogen) in 0.1% Triton X-100/PBS for 1 hr, and incubated with primary antibody (p75 at 1/100 dilution) at 4°C overnight. After washing, the secondary antibody was added, and the sections were incubated for 2 hrs at room temperature. Microscopic images were obtained using a CCD camera (DP72, Olympus, Tokyo) mounted on a fluorescence microscope (BX60, or BX61VS-ASW, Olympus). Cultured cells on coverglasses were fixed in 4% PFA. Antigen retrieval was done by incubation with 100% methanol (-20°C) 10 min, and 0.3% Triton X-100 for 10 min.

     

     

  • 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.

  • 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.

  • 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. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection 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

  • 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.

  • 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:

  • Chen G  et al. Interleukin-1ß Promotes Schwann Cells De-Differentiation in Wallerian Degeneration via the c-JUN/AP-1 Pathway. Front Cell Neurosci 13:304 (2019). Read more (PubMed: 31338026) »
  • Liu Z  et al. p75 neurotrophin receptor regulates NGF-induced myofibroblast differentiation and collagen synthesis through MRTF-A. Exp Cell Res 383:111504 (2019). Read more (PubMed: 31325438) »
See all 20 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Human Cultured Cells (HMC)
Gel Running Conditions
Reduced Non-Denaturing (Native) (4-20% gradient)
Loading amount
40 µg
Specification
HMC
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Oct 27 2019

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

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

Abcam user community

Verified customer

Submitted May 30 2014

Answer

Vielen Dank für Ihre Email.

Gerne send ich Ihnen den Antikörper zu, doch ich bräuchte noch die Umsatzsteuernummer Ihres Institutes, damit Sie nicht unsere Britische Mehrwertsteuer bezahlen müssen...

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

Read More

Answer

Le pido disculpas por la larga espera, he contactado con nuestros colaboradores y mis compañeros del laboratorio para solicitar información acerca de la especificidad de los anticuerpos contra p75NTR.
De las alternativas encontradas, el único que reconocería la región intracelular de la proteína seria ab10494, sintetizado a partir de un péptido sintético correspondientes a la región C terminal (amino ácidos 407-425) de p75NTR de rata.
Respecto a un anticuerpo contra LRP-2, lamentablemente no tenemos información sobre los epítopos que reconocen las distintas alternativas disponibles. Ab52987 se generó a partir de un péptido con secuencia comprendida entre los amino ácidos 350-380.
Siento no poder ser de más ayuda. Sin embargo, le animo a que nos contacte para cualquier otra duda o sugerencia.

Read More

Answer

Thanks for your reply and for the additional information. I agree it does appear that this vial of ab52987 is not working as expected. If you have purchased this antibody within the last 6 months or so I would be happy to offer a replacement, credit or refund. In your reply, please let me know the purchase order number or Abcam order reference number associated with your order so I can process your request.

Read More

Answer

Thank you for your enquiry. I am sorry to hear these two antibodies are giving you trouble. Just a few additional questions regarding your experiments: 1) Have you used a loading control antibody such as an antibody against beta actin to ensure the lysate is of good quality? 2) Have you blotted any other samples with these antibodies? 3) What dilution of primary and secondary was used? 4) How much protein extract was loaded? Thanks for the additional information.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up