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Product Info Summary
| SKU: | M00838 |
|---|---|
| Size: | 100ug |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | IP, IF, WB |
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Product info
Product Name
Anti-HEF1 NEDD9 Monoclonal Antibody
View all NEDD9/CASL/HEF1 Antibodies
SKU/Catalog Number
M00838
Size
100ug
Form
Liquid (sterile filtered)
Description
Boster Bio Anti-HEF1 NEDD9 Monoclonal Antibody (Catalog # M00838). Tested in IF, IP, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store vial at -20°C prior to opening. Aliquot contents and freeze at -20°C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4°C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening. (Ship on dry ice.)
Cite This Product
Anti-HEF1 NEDD9 Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00838)
Host
Mouse
Contents
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01% (w/v) Sodium Azide
Clonality
Monoclonal
Clone Number
Clone: 2G9
Isotype
IgG1 kappa
Immunogen
Anti-HEF1 monoclonal antibody was produced by repeated immunizations with a synthetic peptide corresponding to amino acid residues 82-398 of human HEF1 protein (hHEF1, 843 aa, predicted MW 92.8 kDa).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
M00838 is reactive to NEDD9 in Human, Mouse, Rat
Reconstitution
Observed Molecular Weight
42 kDa
Calculated molecular weight
23510 MW
Background of NEDD9/CASL/HEF1
HEF1, also known as Enhancer of filamentation 1, CRK-associated substrate-related protein, CAS-L, CasL, p105 and Neural precursor cell expressed developmentally down-regulated 9 is the product of the NEDD9 (CASGL) gene. HEF1 functions as a docking protein that plays a central coordinating role for tyrosine-kinase-based signaling related to cell adhesion. HEF1 may also function in transmitting growth control signals between focal adhesions at the cell periphery and the mitotic spindle in response to adhesion or growth factor signals initiating cell proliferation. HEF1 may also play an important role in integrin beta-1 or B cell antigen receptor (BCR) mediated signaling in B- and T-cells. Integrin beta-1 stimulation leads to recruitment of various proteins including CRK, NCK and SHPTP2 to the tyrosine phosphorylated form. HEF1 forms a homodimer and can heterodimerize with HLH proteins ID2, E12, E47 and also with p130cas. HEF1 also forms complexes in vivo with related adhesion focal tyrosine kinase (RAFTK), adapter protein CRKL and LYN kinase and also interacts with MICAL and TXNL4/DIM1. This protein localizes to both the cell nucleus and the cell periphery and is differently localized in fibroblasts and epithelial cells. In fibroblasts, it is predominantly nuclear and in some cells is present in the Golgi apparatus. In epithelial cells, it is localized predominantly in the cell periphery with particular concentration in lamellipodia, but it is also found in the nucleus. HEF1 is widely expressed although higher levels are detected in kidney, lung, and placental tissue. HEF1 is also detected in T-cells, B-cells and diverse cell lines. HEF1 is activated upon induction of cell growth. Cell cycle-regulated processing produces four isoforms: p115, p105, p65, and p55. Isoform p115 arises from p105 phosphorylation and appears later in the cell cycle. Isoform p55 arises from p105 as a result of cleavage at a caspase cleavage-related site and it appears specifically at mitosis. The p65 isoform is poorly detected. Isoforms p105 and p115 are predominantly cytoplasmic and associate with focal adhesions while p55 associates with the mitotic spindle.
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.
Application & Images
Applications
M00838 is guaranteed for IP, IF, 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.
ELISA: 1:5,000 - 1:20,000
IF Microscopy: 1:500
IP: 1:1,000
WB: 1:5,000
Validation Images & Assay Conditions
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Immunofluorescence microscopy using Monoclonal anti-HEFHEF1 localized at focal adhesion sites.The antibody was used at a 1:500 dilution with a 3-sec exposure time. Personal Communication. Elena Pugacheva, Fox Chase Cancer Center, Philadelphia, PA.
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Figure 2. Western blot analysis of NEDD9 using anti-NEDD9 antibody (M00838).
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.
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-NEDD9 antigen affinity purified polyclonal antibody (Catalog # M00838) at 0.5 ug/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.
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Figure 3. Western blot analysis of NEDD9 using anti-NEDD9 antibody (M00838).
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.
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-NEDD9 antigen affinity purified polyclonal antibody (Catalog # M00838) at 0.5 ug/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.
Protein Target Info & Infographic
Gene/Protein Information For NEDD9 (Source: Uniprot.org, NCBI)
Gene Name
NEDD9
Full Name
Enhancer of filamentation 1
Weight
23510 MW
Superfamily
CAS family
Alternative Names
hEF1, NEDD-9, CASL, Cas like docking, CASL, Crk associated substrate related protein, dJ49G10.2, dJ761I2.1, Enhancer of filamentation 1 NEDD9 CAS-L, CAS2, CASL, CASS2, HEF1 neural precursor cell expressed, developmentally down-regulated 9 enhancer of filamentation 1|Cas scaffolding protein family member 2|Crk-associated substrate related protein Cas-L|Enhancer of filamentation 1 p55|cas-like docking|neural precursor cell expressed developmentally down-regulated protein 9|p130Cas-related protein|renal carcinoma NY-REN-12
*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 NEDD9, check out the NEDD9 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 NEDD9: 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-HEF1 NEDD9 Monoclonal Antibody (M00838)
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