Product Info Summary
SKU: | M04157-1 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Mouse |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-eRF1/ETF1 Antibody Picoband™ (monoclonal, 3E5)
SKU/Catalog Number
M04157-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-eRF1/ETF1 Antibody Picoband™ (monoclonal, 3E5) catalog # M04157-1. Tested in 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-eRF1/ETF1 Antibody Picoband™ (monoclonal, 3E5) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M04157-1)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Monoclonal
Clone Number
3 E5
Isotype
Mouse IgG2b
Immunogen
E.coli-derived human eRF1/ETF1 recombinant protein (Position: D9-K342).
*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
M04157-1 is reactive to ETF1 in Human, Mouse, Rat
Applications
M04157-1 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
49 kDa
Calculated molecular weight
49.031kDa
Background of eRF1
Eukaryotic translation termination factor 1 (eRF1), also known asTB3-1, is a protein that in humans is encoded by the ETF1 gene. It is mapped to 5q31.2. This gene encodes a class-1 polypeptide chain release factor. The encoded protein plays an essential role in directing termination of mRNA translation from the termination codons UAA, UAG and UGA. This protein is a component of the SURF complex which promotes degradation of prematurely terminated mRNAs via the mechanism of nonsense-mediated mRNA decay (NMD). Alternate splicing results in multiple transcript variants. Pseudogenes of this gene are found on chromosomes 6, 7, and X.
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), 2-5μg/ml, Human, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human, Mouse, Rat
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (M04157-1).
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 Jurkat whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: human HEPG2 whole cell lysates,
Lane 6: human HEPG2 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 mouse anti-eRF1/ETF1 antigen affinity purified monoclonal antibody (Catalog # M04157-1) 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-mouse 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 # EK1001) with Tanon 5200 system. A specific band was detected for eRF1/ETF1 at approximately 49KD. The expected band size for eRF1/ETF1 is at 49KD.
Click image to see more details
Figure 2. Western blot analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (M04157-1).
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: rat liver tissue lysates,
Lane 2: rat testis tissue lysates,
Lane 3: rat kidney tissue lysates,
Lane 4: rat PC-12 whole cell lysates,
Lane 5: mouse liver tissue lysates,
Lane 6: mouse testis tissue lysates,
Lane 7: mouse kidney tissue lysates,
Lane 8: mouse RAW264.7 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 mouse anti-eRF1/ETF1 antigen affinity purified monoclonal antibody (Catalog # M04157-1) 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-mouse 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 # EK1001) with Tanon 5200 system. A specific band was detected for eRF1/ETF1 at approximately 49KD. The expected band size for eRF1/ETF1 is at 49KD.
Click image to see more details
Figure 3. IHC analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (M04157-1).
eRF1/ETF1 was detected in paraffin-embedded section of human tonsil 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 mouse anti-eRF1/ETF1 Antibody (M04157-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (M04157-1).
eRF1/ETF1 was detected in paraffin-embedded section of human breast 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 mouse anti-eRF1/ETF1 Antibody (M04157-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (M04157-1).
eRF1/ETF1 was detected in paraffin-embedded section of human liver 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 mouse anti-eRF1/ETF1 Antibody (M04157-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (M04157-1).
eRF1/ETF1 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 mouse anti-eRF1/ETF1 Antibody (M04157-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 7. IHC analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (M04157-1).
eRF1/ETF1 was detected in paraffin-embedded section of rat pancreas 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 mouse anti-eRF1/ETF1 Antibody (M04157-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 8. IF analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (M04157-1).
eRF1/ETF1 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 5μg/mL mouse anti-eRF1/ETF1 Antibody (M04157-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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 9. Flow Cytometry analysis of Caco-2 cells using anti-eRF1/ETF1 antibody (M04157-1).
Overlay histogram showing Caco-2 cells stained with M04157-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-eRF1/ETF1 Antibody (M04157-1, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 10. Flow Cytometry analysis of HEPA1-6 cells using anti-eRF1/ETF1 antibody (M04157-1).
Overlay histogram showing HEPA1-6 cells stained with M04157-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-eRF1/ETF1 Antibody (M04157-1, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 11. Flow Cytometry analysis of RH35 cells using anti-eRF1/ETF1 antibody (M04157-1).
Overlay histogram showing RH35 cells stained with M04157-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-eRF1/ETF1 Antibody (M04157-1, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For ETF1 (Source: Uniprot.org, NCBI)
Gene Name
ETF1
Full Name
Eukaryotic peptide chain release factor subunit 1
Weight
49.031kDa
Superfamily
eukaryotic release factor 1 family
Alternative Names
D5S1995; ERF1eRF1; eukaryotic peptide chain release factor subunit 1; Eukaryotic release factor 1; eukaryotic translation termination factor 1; polypeptide chain release factor 1; Protein Cl1; RF1ERF; sup45 (yeast omnipotent suppressor 45) homolog-like 1; SUP45L1; TB3-1MGC111066 ETF1 D5S1995, ERF, ERF1, RF1, SUP45L1, TB3-1 eukaryotic translation termination factor 1 eukaryotic peptide chain release factor subunit 1|polypeptide chain release factor 1|protein Cl1|sup45 (yeast omnipotent suppressor 45) homolog-like 1
*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 ETF1, check out the ETF1 Infographic
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In this infographic, you will see the following information for ETF1: 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-eRF1/ETF1 Antibody Picoband™ (monoclonal, 3E5) (M04157-1)
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