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FCM (Flow Cytometry) (Figure 8. Flow Cytometry analysis of HepG2 cells using anti-ECSIT antibody (AAA19593).Overlay histogram showing HepG2 cells stained with AAA19593 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ECSIT Antibody (AAA19593, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Rabbit anti-Human ECSIT Polyclonal Antibody | anti-ECSIT antibody

Anti-ECSIT Antibody Picoband

Gene Names
ECSIT; SITPEC
Reactivity
Human
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
ECSIT, Antibody; Anti-ECSIT Antibody Picoband; anti-ECSIT antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human
Clonality
Polyclonal
Isotype
Rabbit IgG
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Concentration
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml. (varies by lot)
Applicable Applications for anti-ECSIT antibody
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS), ELISA (EIA)
Application Notes
WB: 0.1-0.25 ug/ml, Human
IHC-P: 2-5 ug/ml, Human
ICC/IF: 5 ug/ml, Human
FC/FACS: 1-3 ug/1x10^6 cells, Human
Direct ELISA: 0.1-0.5 ug/ml, Human
Tested Species: In-house tested species with positive results.
Enhanced Chemiluminescent Kit with anti-Rabbit IgG for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit for IHC(P) and ICC.
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml.
Immunogen
E Coli-derived human ECSIT recombinant protein (Position: E83-S339).
Preparation and Storage
Store at -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months. Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 8. Flow Cytometry analysis of HepG2 cells using anti-ECSIT antibody (AAA19593).Overlay histogram showing HepG2 cells stained with AAA19593 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ECSIT Antibody (AAA19593, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry) (Figure 8. Flow Cytometry analysis of HepG2 cells using anti-ECSIT antibody (AAA19593).Overlay histogram showing HepG2 cells stained with AAA19593 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ECSIT Antibody (AAA19593, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IF (Immunofluorescence)

(Figure 7. IF analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in an immunocytochemical section of PC-3 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 5 ug/mL rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IF (Immunofluorescence) (Figure 7. IF analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in an immunocytochemical section of PC-3 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 5 ug/mL rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IHC (Immunohistchemistry)

(Figure 6. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human lymphadenoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistchemistry) (Figure 6. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human lymphadenoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 5. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human gastric adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 4. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human gastric adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 3. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 3. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 2. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human bladder adenosquamous carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 2. IHC analysis of ECSIT using anti-ECSIT antibody (AAA19593).ECSIT was detected in a paraffin-embedded section of human bladder adenosquamous carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ECSIT Antibody (AAA19593) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of ECSIT using anti-ECSIT antibody (AAA19593).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 30 ug of sample under reducing conditions.Lane 1: human PC-3 whole cell lysates,Lane 2: human Hela whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-ECSIT antigen affinity purified polyclonal antibody (#AAA19593) at 0.25 ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for ECSIT at approximately 49 kDa. The expected band size for ECSIT is at 49 kDa.)

WB (Western Blot) (Figure 1. Western blot analysis of ECSIT using anti-ECSIT antibody (AAA19593).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 30 ug of sample under reducing conditions.Lane 1: human PC-3 whole cell lysates,Lane 2: human Hela whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-ECSIT antigen affinity purified polyclonal antibody (#AAA19593) at 0.25 ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for ECSIT at approximately 49 kDa. The expected band size for ECSIT is at 49 kDa.)
Related Product Information for anti-ECSIT antibody
Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial (ECSIT), also known as SITPEC, is a protein that in humans is encoded by the ECSIT gene. Activation of NF-kB as a result of Toll-like receptor (TLR) and IL-1 receptor signaling is a major component of innate immune responses. Signals from these receptors are relayed by a number of adapter molecules such as TRIF, TIRAP, and MyD88 to kinases such as IRAK and other intermediates such as TNF receptor associated factor (TRAF)-6. ECSIT (evolutionarily conserved signaling intermediate in Toll pathways) was initially identified as a cytoplasmic protein interacting specifically with TNF receptor associated factor (TRAF)-6 in the TLR pathway. Recently however, ECSIT has also been shown to be required for bone morphogenetic protein (Bmp) signaling and mesoderm formation during mouse embryogenesis, indicating the possibility of cross-talk between the TLR/IL-B and Bmp signaling pathways.
Product Categories/Family for anti-ECSIT antibody

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
33,938 Da
NCBI Official Full Name
evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial isoform 2
NCBI Official Synonym Full Names
ECSIT signalling integrator
NCBI Official Symbol
ECSIT
NCBI Official Synonym Symbols
SITPEC
NCBI Protein Information
evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial; ECSIT homolog; likely ortholog of mouse signaling intermediate in Toll pathway evolutionarily conserved
UniProt Protein Name
Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial
UniProt Gene Name
ECSIT
UniProt Entry Name
ECSIT_HUMAN

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Product Notes

The ECSIT ecsit (Catalog #AAA19593) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-ECSIT Antibody Picoband reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's ECSIT can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS), ELISA (EIA). WB: 0.1-0.25 ug/ml, Human IHC-P: 2-5 ug/ml, Human ICC/IF: 5 ug/ml, Human FC/FACS: 1-3 ug/1x10^6 cells, Human Direct ELISA: 0.1-0.5 ug/ml, Human Tested Species: In-house tested species with positive results. Enhanced Chemiluminescent Kit with anti-Rabbit IgG for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit for IHC(P) and ICC. Researchers should empirically determine the suitability of the ECSIT ecsit for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "ECSIT, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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