Product Info Summary
SKU: | A05258-3 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-AFAP/AFAP1 Antibody Picoband™
SKU/Catalog Number
A05258-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-AFAP/AFAP1 Antibody Picoband™ catalog # A05258-3. Tested in ELISA, Flow Cytometry, IHC, 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-AFAP/AFAP1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05258-3)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human AFAP/AFAP1 recombinant protein (Position: A498-T730).
*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
A05258-3 is reactive to AFAP1 in Human, Mouse, Rat
Applications
A05258-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Observed Molecular Weight
120 kDa
Calculated molecular weight
80.725kDa
Background of AFAP
Actin filament-associated protein 1 is a protein that in humans is encoded by the AFAP1 gene. The protein encoded by this gene is a Src binding partner. It may represent a potential modulator of actin filament integrity in response to cellular signals, and may function as an adaptor protein by linking Src family members and/or other signaling proteins to actin filaments. Multiple transcript variants encoding different isoforms have been found for this gene.
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, 0.1-0.25μg/ml,Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/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 AFAP/AFAP1 using anti-AFAP/AFAP1 antibody (A05258-3).
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 Hela whole cell lysates,
Lane 2: human SH-SY5Y whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: human U-87MG whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat lung tissue lysates,
Lane 7: rat stomach tissue lysates,
Lane 8: mouse brain tissue lysates,
Lane 9: mouse lung tissue lysates,
Lane 10: mouse stomach tissue 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-AFAP/AFAP1 antigen affinity purified polyclonal antibody (Catalog # A05258-3) at 0.25 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 AFAP/AFAP1 at approximately 120KD. The expected band size for AFAP/AFAP1 is at 120KD.
Click image to see more details
Figure 2. Flow Cytometry analysis of A549 cells using anti-AFAP/AFAP1 antibody (A05258-3).
Overlay histogram showing A549 cells stained with A05258-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AFAP/AFAP1 Antibody (A05258-3, 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.
Click image to see more details
Figure 3. Flow Cytometry analysis of U20S cells using anti-AFAP/AFAP1 antibody (A05258-3).
Overlay histogram showing U20S cells stained with A05258-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AFAP/AFAP1 Antibody (A05258-3, 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.
Click image to see more details
Figure 4. IHC analysis of AFAP/AFAP1 using anti-AFAP/AFAP1 antibody (A05258-3).
AFAP/AFAP1 was detected in paraffin-embedded section of human lung 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 2μg/ml rabbit anti-AFAP/AFAP1 Antibody (A05258-3) 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.
Click image to see more details
Figure 5. IHC analysis of AFAP/AFAP1 using anti-AFAP/AFAP1 antibody (A05258-3).
AFAP/AFAP1 was detected in paraffin-embedded section of human prostatic 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 2μg/ml rabbit anti-AFAP/AFAP1 Antibody (A05258-3) 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.
Protein Target Info & Infographic
Gene/Protein Information For AFAP1 (Source: Uniprot.org, NCBI)
Gene Name
AFAP1
Full Name
Actin filament-associated protein 1
Weight
80.725kDa
Alternative Names
actin filament associated protein 1; AFAP; AFAP1; AFAP-110; FLJ56849 AFAP1 AFAP, AFAP-11010, AFAP1 actin filament associated protein 1 actin filament-associated protein 1|110 kDa actin filament-associated protein|actin filament-associated protein, 110 kDa
*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 AFAP1, check out the AFAP1 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 AFAP1: 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-AFAP/AFAP1 Antibody Picoband™ (A05258-3)
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