Product Info Summary
SKU: | A04649 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Agrin/AGRN Picoband™ Antibody
SKU/Catalog Number
A04649
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Agrin/AGRN Picoband™ Antibody catalog # A04649. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store 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 freeze-thaw cycles.
Cite This Product
Anti-Agrin/AGRN Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04649)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Agrin/AGRN recombinant protein (Position: S1864-P2068).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A04649 is reactive to Agrn in Human, Mouse, Rat
Applications
A04649 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
205-217 kDa
Calculated molecular weight
207.539kDa
Background of Agrin
Agrin is a protein that in humans is encoded by the AGRN gene. It is mapped to 1p36.33. This gene encodes one of several proteins that are critical in the development of the neuromuscular junction (NMJ), as identified in mouse knock-out studies. The encoded protein contains several laminin G, Kazal type serine protease inhibitor, and epidermal growth factor domains. Additional post-translational modifications occur to add glycosaminoglycans and disulfide bonds. In one family with congenital myasthenic syndrome affecting limb-girdle muscles, a mutation in this gene was found. Alternative splicing results in multiple transcript variants encoding different isoforms.
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.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
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, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of AGRN using anti-AGRN antibody (A04649).
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 30ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: rat lung tissue lysates,
Lane 3: mouse HEPA1-6 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-AGRN antigen affinity purified polyclonal antibody (Catalog # A04649) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for AGRN at approximately 205-217KD. The expected band size for AGRN is at 217KD.
Click image to see more details
Figure 2. IHC analysis of Agrin/AGRN using anti-Agrin/AGRN antibody (A04649).
Agrin/AGRN was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Agrin/AGRN Antibody (A04649) 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.
Click image to see more details
Figure 3. IF analysis of Agrin/AGRN using anti-Agrin/AGRN antibody (A04649).
Agrin/AGRN was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-Agrin/AGRN Antibody (A04649) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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.
Click image to see more details
Figure 4. Flow Cytometry analysis of A549 cells using anti-Agrin/AGRN antibody (A04649).
Overlay histogram showing A549 cells stained with A04649 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Agrin/AGRN Antibody (A04649, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For Agrn (Source: Uniprot.org, NCBI)
Gene Name
Agrn
Full Name
Agrin
Weight
207.539kDa
Alternative Names
agrin proteoglycan; Agrin; AGRN Agrn|A, Agrin, NMF38, nmf380|agrin|agrin
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on Agrn, check out the Agrn Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for Agrn: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Agrin/AGRN Picoband™ Antibody (A04649)
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