Recombinant
RabMAb

Recombinant Anti-p75 NGF Receptor antibody [EP1039Y] - Low endotoxin, Azide free (ab221212)

Overview

  • Product name

    Anti-p75 NGF Receptor antibody [EP1039Y] - Low endotoxin, Azide free
    See all p75 NGF Receptor primary antibodies
  • Description

    Rabbit monoclonal [EP1039Y] to p75 NGF Receptor - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide aa 350-450. The exact sequence is proprietary. This protein is only 427 aa in length

  • Positive control

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

    ab221212 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab221212 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 Use at an assay dependent concentration. Detects a band of approximately 75 kDa (predicted molecular weight: 45 kDa).
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

  • 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 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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).

References

This product has been referenced in:

See all 8 Publications for this product

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