SKU A00081-3
Size 100μg/vial
Reactivity Human
Clonality Polyclonal
Host Rabbit
Applications IHC-P, WB

Overview

Product Name Anti-APP/C99 Picoband™ Antibody
SKU/Catalog Number A00081-3
Storage & Handling At -20°C; for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for APP/C99 detection. Tested with WB, IHC-P in Human.
Cite This Product Anti-APP/C99 Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00081-3)
Host Rabbit
Contents/Buffer Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence of human APP/C99(DAEFRHDSGYEVHHQKLVFFAEDVGSNK).
Reactivity Human

Assay Details

Assay Dilutions Overview

Concentration: 0.5-1mg/ml, actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
Western blot, 0.1-0.5μg/ml
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).

Images And Assay Conditions

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Figure 1. Western blot analysis of APP/C99 using anti-APP/C99 antibody (A00081-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human T-47D whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-APP/C99 antigen affinity purified polyclonal antibody (Catalog # A00081-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for APP/C99 at approximately 99KD. The expected band size for APP/C99 is at 87KD.

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Figure 2. IHC analysis of C99 using anti-C99 antibody (A00081-3).
C99 was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-C99 Antibody (A00081-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of C99 using anti-C99 antibody (A00081-3).
C99 was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-C99 Antibody (A00081-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

/A/0/A00081 3 C99 primary antibodies IHC testing 4.jpg

Figure 4. IHC analysis of C99 using anti-C99 antibody (A00081-3).
C99 was detected in paraffin-embedded section of human renal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-C99 Antibody (A00081-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P05067
Gene Name APP
Protein Name amyloid beta precursor protein
Tissue Specificity Expressed in all fetal tissues examined with highest levels in brain, kidney, heart and spleen. Weak expression in liver. In adult brain, highest expression found in the frontal lobe of the cortex and in the anterior perisylvian cortex-opercular gyri. Moderate expression in the cerebellar cortex, the posterior perisylvian cortex-opercular gyri and the temporal associated cortex. Weak expression found in the striate, extra-striate and motor cortices. Expressed in cerebrospinal fluid, and plasma. Isoform APP695 is the predominant form in neuronal tissue, isoform APP751 and isoform APP770 are widely expressed in non-neuronal cells. Isoform APP751 is the most abundant form in T-lymphocytes. Appican is expressed in astrocytes.
Alternative Names Amyloid-beta A4 protein; ABPP; APPI; APP; Alzheimer disease amyloid protein; Amyloid precursor protein; Amyloid-beta precursor protein; Cerebral vascular amyloid peptide; CVAP; PreA4; Protease nexin-II; PN-II; S-APP-alpha; S-APP-beta; Beta-secretase C-terminal fragment; Beta-CTF; Abeta42; Beta-APP42; Abeta40; Beta-APP40; Alpha-secretase C-terminal fragment; Alpha-CTF; Amyloid intracellular domain 59; AICD-59; AID(59); Gamma-CTF(59); Amyloid intracellular domain 57; AICD-57; AID(57); Gamma-CTF(57); Amyloid intracellular domain 50; AICD-50; AID(50); Gamma-CTF(50); APP; A4; AD1

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Functions as a cell surface receptor and performs physiological functions on the surface of neurons relevant to neurite growth, neuronal adhesion and axonogenesis. Involved in cell mobility and transcription regulation through protein-protein interactions. Can promote transcription activation through binding to APBB1-KAT5 and inhibits Notch signaling through interaction with Numb. Couples to apoptosis-inducing pathways such as those mediated by G(O) and JIP. Inhibits G(o) alpha ATPase activity. Acts as a kinesin I membrane receptor, mediating the axonal transport of beta-secretase and presenilin 1. Involved in copper homeostasis/oxidative stress through copper ion reduction. In vitro, copper-metallated APP induces neuronal death directly or is potentiated through Cu2+-mediated low-density lipoprotein oxidation. Can regulate neurite outgrowth through binding to components of the extracellular matrix such as heparin and collagen I and IV. The splice isoforms that contain the BPTI domain possess protease inhibitor activity. Induces a AGER-dependent pathway that involves activation of p38 MAPK, resulting in internalization of amyloid-beta peptide and leading to mitochondrial dysfunction in cultured cortical neurons. Provides Cu2+ ions for GPC1 which are required for release of nitric oxide (NO) and subsequent degradation of the heparan sulfate chains on GPC1.
Research Areas Adapters, Apoptosis, Associated Proteins, Cell Biology, Cytoplasmic, Developmental Biology, Intracellular, Nervous System Development, Neural Signal Transduction, Neurogenesis, Neurology Process, Neuroscience, Organelle Proteins, Organogenesis, Protein Trafficking, Signal Transduction

*You can search these to find other products in these research areas.
Background Amyloid precursor protein (APP) is an integral membrane protein expressed in many tissues and concentrated in the synapses of neurons. It is mapped to 21q21.3. This gene encodes a cell surface receptor and transmembrane precursor protein that is cleaved by secretases to form a number of peptides. Some of these peptides are secreted and can bind to the acetyltransferase complex APBB1/TIP60 to promote transcriptional activation, while others form the protein basis of the amyloid plaques found in the brains of patients with Alzheimer disease. In addition, two of the peptides are antimicrobial peptides, having been shown to have bacteriocidal and antifungal activities. Mutations in this gene have been implicated in autosomal dominant Alzheimer disease and cerebroarterial amyloidosis (cerebral amyloid angiopathy). Multiple transcript variants encoding several different isoforms have been found for this gene.

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