Anti-Entactin/NID1 Antibody Picoband®
PRODUCTS SPECIFICATIONS
DESCRIPTION
Product Name
Anti-Entactin/NID1 Antibody Picoband®
SKU/Catalog Number
A05621
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Entactin/NID1 Antibody Picoband® catalog # A05621. Tested in WB, IHC, IF, Flow Cytometry, ELISA applications. This antibody reacts with Human, Mouse. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months.Avoid repeated freezing and thawing.
Cite This Product
Anti-Entactin/NID1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05621)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Immunogen
E.coli-derived human Entactin/NID1 recombinant protein (Position: R31-H1209). Human Entactin/NID1 shares 85.6% amino acid (aa) sequence identity with mouse Entactin/NID1.
Reactive Species
A05621 is reactive to NID1 in Human, Mouse
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Observed Molecular Weight
150 kDa
Calculated molecular weight
136.4 kDa
Background of NID1
This gene encodes a member of the nidogen family of basement membrane glycoproteins. The protein interacts with several other components of basement membranes, and may play a role in cell interactions with the extracellular matrix.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Applications
A05621 is guaranteed for ELISA, Flow Cytometry, IF, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 μg/ml, Human, Mouse
Immunohistochemistry, 2-5 μg/ml, Human
Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml
Validation Images & Assay Conditions
Western blot analysis of NID1 using anti-NID1 antibody (A05621).
Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human THP-1 whole cell lysates,
Lane 3: human 293T whole cell lysates,
Lane 4: mouse heart tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-NID1 antigen affinity purified polyclonal antibody (A05621) 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 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for NID1 at approximately 150 kDa. The expected band size for NID1 is at 136 kDa.
IHC analysis of NID1 using anti-NID1 antibody (A05621).
NID1 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NID1 Antibody (A05621) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
IF analysis of NID1 using anti-NID1 antibody (A05621).
NID1 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-NID1 Antibody (A05621) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of HepG2 cells using anti-NID1 antibody (A05621).
Overlay histogram showing HepG2 cells stained with A05621 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-NID1 Antibody (A05621, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
