Recombinant Anti-p75 NGF Receptor antibody [EP1039Y] - Low endotoxin, Azide free (ab221212)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1039Y] to p75 NGF Receptor - Low endotoxin, Azide free
- Suitable for: IP, IHC-P, ICC/IF, WB, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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 -
Specificity
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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Tested applications
Suitable for: IP, IHC-P, ICC/IF, WB, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- PC12 cell membrane lysate, PC-12 cell lysate Human brain gilioma tissue
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General notes
ab221212 is the carrier-free version of ab52987.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Dissociation constant (KD)
KD = 3.25 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1039Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-p75 NGF Receptor antibody [EP1039Y] (ab195179)
- Alexa Fluor® 647 Anti-p75 NGF Receptor antibody [EP1039Y] (ab195180)
- APC Anti-p75 NGF Receptor antibody [EP1039Y] (ab224996)
- PE Anti-p75 NGF Receptor antibody [EP1039Y] (ab310935)
- Alexa Fluor® 594 Anti-p75 NGF Receptor antibody [EP1039Y] (ab311687)
- Alexa Fluor® 568 Anti-p75 NGF Receptor antibody [EP1039Y] (ab312963)
- Alexa Fluor® 555 Anti-p75 NGF Receptor antibody [EP1039Y] (ab313172)
- Anti-p75 NGF Receptor antibody [EP1039Y] (ab52987)
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
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 |
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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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
modificationsN- 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
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Database links
- Entrez Gene: 4804 Human
- Entrez Gene: 18053 Mouse
- Entrez Gene: 24596 Rat
- Omim: 162010 Human
- SwissProt: P08138 Human
- SwissProt: Q9Z0W1 Mouse
- SwissProt: P07174 Rat
- Unigene: 415768 Human
see all -
Alternative names
- CD271 antibody
- CD271 antigen antibody
- Gp80 LNGFR antibody
see all
Images
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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).
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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).
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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).
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Intracellular 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).
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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).
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Intracellular 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).
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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).
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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).
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52987).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (10)
ab221212 has been referenced in 10 publications.
- Zagozewski J et al. Combined MEK and JAK/STAT3 pathway inhibition effectively decreases SHH medulloblastoma tumor progression. Commun Biol 5:697 (2022). PubMed: 35835937
- Zierold S et al. Brain-Derived Neurotrophic Factor Expression and Signaling in Different Perivascular Adipose Tissue Depots of Patients With Coronary Artery Disease. J Am Heart Assoc 10:e018322 (2021). PubMed: 33666096
- Pascal D et al. Characterization of Glial Cell Models and In Vitro Manipulation of the Neuregulin1/ErbB System. Biomed Res Int 2014:310215 (2014). WB ; Rat . PubMed: 25177687
- Lumb R et al. Neuropilins define distinct populations of neural crest cells. Neural Dev 9:24 (2014). PubMed: 25363691
- Kato H et al. Hypoxia induces an undifferentiated phenotype of oral keratinocytes in vitro. Cells Tissues Organs 199:393-404 (2014). WB ; Human . PubMed: 25720390
- Madelung A et al. A novel immunohistochemical sequential multi-labelling and erasing technique enables epitope characterization of bone marrow pericytes in primary myelofibrosis. Histopathology 60:554-60 (2012). IHC-P ; Human . PubMed: 22250648
- Liu Q et al. Human neural crest stem cells derived from human ESCs and induced pluripotent stem cells: induction, maintenance, and differentiation into functional schwann cells. Stem Cells Transl Med 1:266-78 (2012). ICC/IF ; Human . PubMed: 23197806
- Li Y et al. Accumulation of nerve growth factor and its receptors in the uterus and dorsal root ganglia in a mouse model of adenomyosis. Reprod Biol Endocrinol 9:30 (2011). WB, IHC-P ; Mouse . PubMed: 21385399
- Zabierowski SE et al. Direct reprogramming of melanocytes to neural crest stem-like cells by one defined factor. Stem Cells 29:1752-62 (2011). ICC/IF . PubMed: 21948558
- Li L et al. Human dermal stem cells differentiate into functional epidermal melanocytes. J Cell Sci 123:853-60 (2010). ICC/IF ; Human . PubMed: 20159965