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Product Info Summary
| SKU: | A04576-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Monkey, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-EHD3 Antibody Picoband™
SKU/Catalog Number
A04576-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-EHD3 Antibody Picoband™ catalog # A04576-1. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Monkey, 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-EHD3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04576-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human EHD3, which shares 93.8% and 100% amino acid (aa) sequence identity with mouse and rat EHD3, respectively.
*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
A04576-1 is reactive to EHD3 in Human, Monkey, Mouse, Rat
Applications
A04576-1 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
70 kDa
Calculated molecular weight
Background of EHD3
Eps15 homology domain-containing protein 3, abbreviated as EDH3 and also known as PAST3, is a protein encoded by the EHD3 gene. The Eps15 homology (EH) domain-containing protein family consists of four members, EHD1, EHD2, EHD3, and EHD4. The chromosomal locations of the human EHD genes are as follows: EHD1 maps to 11q13, EHD2 maps to 19q13.3, EHD3 maps to 2p21, and EHD4 maps to 15q11.1 (1-3). The encoded proteins of all EHD family members contain multiple conserved regions, which include an amino-terminal nucleotide-binding consensus site, a bipartite nuclear localization signal, and a carboxy-terminal EH protein-binding domain with an EF-hand motif (3,4). EHD1 is ubiquitously expressed with increased expression in testis (2,3). EHD2, EHD3, and EHD4 have more specific expression with EHD2 highly expressed in heart, EHD3 expressed in brain, kidney, liver, placenta, ovary, and heart, and EHD4 expressed in heart, placenta, and pancreas (1,5,6). The EHD proteins may participate in ligand-induced endocytosis (3-5).
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.
Assay dilution & Images
Reconstitution
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, Monkey, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of EHD3 using anti-EHD3 antibody (A04576-1).
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 30ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human CCRF-CEM whole cell lysates,
Lane 5: human U-87MG whole cell lysates,
Lane 6: human SY-SY5Y whole cell lysates,
Lane 7: monkey heart 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-EHD3 antigen affinity purified polyclonal antibody (Catalog # A04576-1) 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 EHD3 at approximately 70KD. The expected band size for EHD3 is at 70KD.
Click image to see more details
Figure 10. IHC analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 was detected in paraffin-embedded section of mouse brain 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-EHD3 Antibody (A04576-1) 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 2. Western blot analysis of EHD3 using anti-EHD3 antibody (A04576-1).
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 30ug of sample under reducing conditions.
Lane 1: rat heart tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat kidney tissue lysates,
Lane 4: rat liver tissue lysates,
Lane 5: mouse heart tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse kidney tissue lysates,
Lane 8: mouse liver 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-EHD3 antigen affinity purified polyclonal antibody (Catalog # A04576-1) 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 EHD3 at approximately 70KD. The expected band size for EHD3 is at 70KD.
Click image to see more details
Figure 3. IHC analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 was detected in paraffin-embedded section of human ovarian serous adenocarcinoma 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-EHD3 Antibody (A04576-1) 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 4. IHC analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 was detected in paraffin-embedded section of human ovarian serous adenocarcinoma 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-EHD3 Antibody (A04576-1) 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 EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 was detected in paraffin-embedded section of human renal clear cell carcinoma 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-EHD3 Antibody (A04576-1) 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 6. IHC analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 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 rabbit anti-EHD3 Antibody (A04576-1) 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 7. IHC analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 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-EHD3 Antibody (A04576-1) 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 8. IHC analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 was detected in paraffin-embedded section of human rectal 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-EHD3 Antibody (A04576-1) 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 9. IHC analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 was detected in paraffin-embedded section of human gastric adenocarcinoma 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-EHD3 Antibody (A04576-1) 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 11. IHC analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 was detected in paraffin-embedded section of rat brain 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-EHD3 Antibody (A04576-1) 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 12. IF analysis of EHD3 using anti-EHD3 antibody (A04576-1).
EHD3 was detected in immunocytochemical section of MCF-7 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 rabbit anti-EHD3 Antibody (A04576-1) 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 13. Flow Cytometry analysis of MCF-7 cells using anti-EHD3 antibody (A04576-1).
Overlay histogram showing MCF-7 cells stained with A04576-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-EHD3 Antibody (A04576-1, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For EHD3 (Source: Uniprot.org, NCBI)
Gene Name
EHD3
Full Name
EH domain-containing protein 3
Weight
Superfamily
TRAFAC class dynamin-like GTPase superfamily
Alternative Names
EH domain containing 3; EH domain-containing protein 3; EHD2; EH-domain containing 3; PAST homolog 3; PAST3 EHD3 PAST3 EH domain containing 3 EH domain-containing protein 3|PAST homolog 3
*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 EHD3, check out the EHD3 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 EHD3: 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-EHD3 Antibody Picoband™ (A04576-1)
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