Testing Data
(Image: Western blot analysis of extracts from various cell types using TCF4 (C48H11) Rabbit mAb.)
TCF4 (C48H11) Antibody | anti-TCF4 antibody
TCF4 (C48H11), Monoclonal Antibody, Rabbit
Gene Names
TCF4; E2-2; ITF2; PTHS; SEF2; FECD3; ITF-2; SEF-2; TCF-4; SEF2-1; SEF2-1A; SEF2-1B; SEF2-1D; bHLHb19
Reactivity
Chicken, Human, Mouse, Rat
Applications
Western Blot, Immunoprecipitation
Synonyms
TCF4 (C48H11); Monoclonal Antibody; Rabbit; anti-TCF4 antibody
Reactivity
Chicken, Human, Mouse, Rat
Specificity
TCF4 (C48H11) Rabbit mAb detects endogenous levels of total TCF4 protein.
Form/Format
Liquid in 10mM sodium HEPES (pH7.5), 150mM NaCl, 100ug BSA and 50% glycerol.
Sequence Length
773
Applicable Applications for anti-TCF4 antibody
Western Blot (WB), Immunoprecipitation (IP)
Application Notes
Western Blot: 1:1000
Immunoprecipitation: 1:50
Dilutions listed only as a recommendation.
Optimal dilution should be determined by investigator.
Western Blot: Sample Preparation
1. Treat cells by adding fresh media containing regulator for desired time.
2. Aspirate media from cultures; wash cells with 1X PBS; aspirate.
3. Lyse cells by adding 1X SDS sample buffer (100 ul per well of 6-well plate or 500 ul per plate of 10 cmplate). Immediately scrape cells from plate and transfer the extract to a microcentrifuge tube. Keep on ice.
4. Sonicate for 10–15 seconds to shear DNA and reduce sample viscosity.
5. Heat a 20 ul sample to 95–100°C for 5 minutes; cool on ice.
6. Microcentrifuge for 5 minutes.
7. Load 20 ul onto SDS-PAGE gel (10 cm x 10 cm).
8. Electrotransfer to nitrocellulose (or PVDF) membrane.
Membrane Blocking and Antibody Incubations
Note: Volumes for 10 cm x 10 cm (100 cm2) membrane; for different sized membranes, adjust vol. accordingly.
1. (Optional) After transfer, wash nitrocellulose membrane with 25 ml TBS for 5 minutes at room temperature.
2. Incubate membrane in 25 ml of blocking buffer for 1 hour at room temperature.
3. Wash three times for 5 minutes each with 15 ml of TBS/T.
4. Incubate membrane and primary antibody (at the appropriate dilution) in 10 ml primary antibody dilutionbuffer with gentle agitation overnight at 4°C.
5. Wash three times for 5 minutes each with 15 ml of TBS/T.
6. Incubate membrane with HRP-conjugated secondary antibody (1:2000) in 10 ml of blocking buffer withgentle agitation for 1 hour at room temperature.
7. Wash three times for 5 minutes each with 15 ml of TBS/T.
8. Process membranes using enhanced chemiluminescence.
Immunoprecipitation:
Preparing Cell Lysates
1. Aspirate media. Treat cells by adding fresh media containing regulator fordesired time.
2. To harvest cells under nondenaturing conditions, remove media and rinse cells once with ice-cold PBS.
3. Remove PBS and add 0.5 ml ice-cold 1X cell lysis buffer to each plate (10 cm)and incubate the plates on ice for 5 minutes.
4. Scrape cells off the plates and transfer to microcentrifuge tubes. Keep on ice.
5. Sonicate samples on ice three times for 5 seconds each.
6.Microcentrifuge for 10 minutes at 14,000 X g, 4°C, and transfer the supernatantto a new tube. If necessary, lysate can be stored at –80°C.
Immunoprecipitation
Optional: It may be necessary to perform a lysate pre-cleaning step to reduce non-specificbinding to the Protein A/G agarose beads (See section below).
1. Take 200 ul cell lysate and add primary antibody. Incubate with gentle rockingovernight at 4°C.
2. Add either protein A or G agarose beads (20 ul of 50% bead slurry). Incubate with gentle rocking for 1–3 hours at 4°C.
3. Microcentrifuge for 30 seconds at 4°C. Wash pellet five times with 500 ul of 1Xcell lysis buffer. Keep on ice during washes.
4. Resuspend the pellet with 20 ul 3X SDS sample buffer. Vortex, then microcentrifugefor 30 seconds.
5. Heat the sample to 95–100°C for 2–5 minutes and microcentrifuge for 1 minuteat 14,000 X g.
6. Load the sample (15–30 ul) on SDS-PAGE gel (12–15%).
7. Analyze sample by Western blotting (see Western Immunoblotting Protocol).
Cell Lysate Pre-Cleaning (Optional)
1. Take 200 ul cell lysate and add to either Protein A or G agarose beads (20 ul of50% bead slurry).
2. Incubate at 4°C for 30 – 60 minutes.
3. Spin for 10 minutes at 4°C. Transfer the supernatant to a fresh tube.
4. Proceed to step 1 of Immunoprecipitation.
Immunoprecipitation: 1:50
Dilutions listed only as a recommendation.
Optimal dilution should be determined by investigator.
Western Blot: Sample Preparation
1. Treat cells by adding fresh media containing regulator for desired time.
2. Aspirate media from cultures; wash cells with 1X PBS; aspirate.
3. Lyse cells by adding 1X SDS sample buffer (100 ul per well of 6-well plate or 500 ul per plate of 10 cmplate). Immediately scrape cells from plate and transfer the extract to a microcentrifuge tube. Keep on ice.
4. Sonicate for 10–15 seconds to shear DNA and reduce sample viscosity.
5. Heat a 20 ul sample to 95–100°C for 5 minutes; cool on ice.
6. Microcentrifuge for 5 minutes.
7. Load 20 ul onto SDS-PAGE gel (10 cm x 10 cm).
8. Electrotransfer to nitrocellulose (or PVDF) membrane.
Membrane Blocking and Antibody Incubations
Note: Volumes for 10 cm x 10 cm (100 cm2) membrane; for different sized membranes, adjust vol. accordingly.
1. (Optional) After transfer, wash nitrocellulose membrane with 25 ml TBS for 5 minutes at room temperature.
2. Incubate membrane in 25 ml of blocking buffer for 1 hour at room temperature.
3. Wash three times for 5 minutes each with 15 ml of TBS/T.
4. Incubate membrane and primary antibody (at the appropriate dilution) in 10 ml primary antibody dilutionbuffer with gentle agitation overnight at 4°C.
5. Wash three times for 5 minutes each with 15 ml of TBS/T.
6. Incubate membrane with HRP-conjugated secondary antibody (1:2000) in 10 ml of blocking buffer withgentle agitation for 1 hour at room temperature.
7. Wash three times for 5 minutes each with 15 ml of TBS/T.
8. Process membranes using enhanced chemiluminescence.
Immunoprecipitation:
Preparing Cell Lysates
1. Aspirate media. Treat cells by adding fresh media containing regulator fordesired time.
2. To harvest cells under nondenaturing conditions, remove media and rinse cells once with ice-cold PBS.
3. Remove PBS and add 0.5 ml ice-cold 1X cell lysis buffer to each plate (10 cm)and incubate the plates on ice for 5 minutes.
4. Scrape cells off the plates and transfer to microcentrifuge tubes. Keep on ice.
5. Sonicate samples on ice three times for 5 seconds each.
6.Microcentrifuge for 10 minutes at 14,000 X g, 4°C, and transfer the supernatantto a new tube. If necessary, lysate can be stored at –80°C.
Immunoprecipitation
Optional: It may be necessary to perform a lysate pre-cleaning step to reduce non-specificbinding to the Protein A/G agarose beads (See section below).
1. Take 200 ul cell lysate and add primary antibody. Incubate with gentle rockingovernight at 4°C.
2. Add either protein A or G agarose beads (20 ul of 50% bead slurry). Incubate with gentle rocking for 1–3 hours at 4°C.
3. Microcentrifuge for 30 seconds at 4°C. Wash pellet five times with 500 ul of 1Xcell lysis buffer. Keep on ice during washes.
4. Resuspend the pellet with 20 ul 3X SDS sample buffer. Vortex, then microcentrifugefor 30 seconds.
5. Heat the sample to 95–100°C for 2–5 minutes and microcentrifuge for 1 minuteat 14,000 X g.
6. Load the sample (15–30 ul) on SDS-PAGE gel (12–15%).
7. Analyze sample by Western blotting (see Western Immunoblotting Protocol).
Cell Lysate Pre-Cleaning (Optional)
1. Take 200 ul cell lysate and add to either Protein A or G agarose beads (20 ul of50% bead slurry).
2. Incubate at 4°C for 30 – 60 minutes.
3. Spin for 10 minutes at 4°C. Transfer the supernatant to a fresh tube.
4. Proceed to step 1 of Immunoprecipitation.
Type Info
Rabbit IgG
Immunogen
Synthetic peptide (KLH-coupled) corresponding to (KLH-coupled) corresponding to Leu330 of human TCF4.
Preparation and Storage
Storeat -20ºC. Do not aliquot the antibody. Stable for up to at least 6 months. Repeated freeze/thaw cycles compromise the integrity of the antiserum.
Testing Data
(Image: Western blot analysis of extracts from various cell types using TCF4 (C48H11) Rabbit mAb.)
Testing Data
(Image: Western blot analysis of extracts from various cell types using TCF4 (C48H11) Rabbit mAb.)
Related Product Information for anti-TCF4 antibody
LEF1 and TCF are members of the high mobility group (HMG) DNA binding protein family of transcription factors which consists of the following: Lymphoid enhancer factor 1 (LEF1), T Cell Factor 1 (TCF1), TCF3 and TCF4 (1). LEF1 and TCF1 were originally identified as important factors regulating early lymphoid development (2) and act downstream in Wnt signaling. LEF1/TCF bind to Wnt response elements to provide a docking site for b-catenin, which translocates to the nucleus to promote the transcription of target genes upon activation of Wnt signaling (3). LEF1/TCF proteins are dynamically expressed during development and aberrant activation of the Wnt signaling pathway is involved in many types of cancers including colon cancer (4,5).
TCF4, also known as TCF7L2, is expressed widely during development. Gene targeting studies indicate that TCF4 is required to maintain the crypt stem cells of the small intestine (6,7). TCF4 has several splicing isoforms that are expressed differentially in tissues and during cancer progression (8,9). Studies also indicate that a variant of the TCF4 gene confers an increased risk of type 2 diabetes (10).
TCF4, also known as TCF7L2, is expressed widely during development. Gene targeting studies indicate that TCF4 is required to maintain the crypt stem cells of the small intestine (6,7). TCF4 has several splicing isoforms that are expressed differentially in tissues and during cancer progression (8,9). Studies also indicate that a variant of the TCF4 gene confers an increased risk of type 2 diabetes (10).
Product Categories/Family for anti-TCF4 antibody
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
NCBI Official Full Name
transcription factor 4 isoform c
NCBI Official Synonym Full Names
transcription factor 4
NCBI Official Symbol
TCF4
NCBI Official Synonym Symbols
E2-2; ITF2; PTHS; SEF2; FECD3; ITF-2; SEF-2; TCF-4; SEF2-1; SEF2-1A; SEF2-1B; SEF2-1D; bHLHb19
NCBI Protein Information
transcription factor 4
UniProt Protein Name
Transcription factor 4
Protein Family
UniProt Gene Name
TCF4
UniProt Synonym Gene Names
BHLHB19; ITF2; SEF2; TCF-4; bHLHb19; ITF-2; SEF-2
UniProt Entry Name
ITF2_HUMAN
NCBI Description
This gene encodes transcription factor 4, a basic helix-loop-helix transcription factor. The encoded protein recognizes an Ephrussi-box ('E-box') binding site ('CANNTG') - a motif first identified in immunoglobulin enhancers. This gene is broadly expressed, and may play an important role in nervous system development. Defects in this gene are a cause of Pitt-Hopkins syndrome. Multiple alternatively spliced transcript variants that encode different proteins have been described. [provided by RefSeq, Aug 2011]
Uniprot Description
TCF4: Transcription factor that binds to the immunoglobulin enchancer Mu-E5/KE5-motif. Involved in the initiation of neuronal differentiation. Activates transcription by binding to the E box (5'-CANNTG-3'). Binds to the E-box present in the somatostatin receptor 2 initiator element (SSTR2-INR) to activate transcription. Preferentially binds to either 5'-ACANNTGT-3' or 5'-CCANNTGG-3'. Efficient DNA binding requires dimerization with another bHLH protein. Forms homo- or heterooligomers with myogenin. Interacts with HIVEP2. Interacts with NEUROD2. Expressed in adult heart, brain, placenta, skeletal muscle and to a lesser extent in the lung. In developing embryonic tissues, expression mostly occurs in the brain. 3 isoforms of the human protein are produced by alternative splicing.
Protein type: DNA-binding; Transcription factor
Chromosomal Location of Human Ortholog: 18q21.1
Cellular Component: transcription factor complex; nucleus
Molecular Function: protein C-terminus binding; protein binding; DNA binding; protein heterodimerization activity; transcription factor activity
Biological Process: transcription initiation from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; transcription initiation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of neuron differentiation; protein-DNA complex assembly
Disease: Corneal Dystrophy, Fuchs Endothelial, 3; Pitt-hopkins Syndrome
Research Articles on TCF4
Similar Products
Product Notes
The TCF4 tcf4 (Catalog #AAA555278) is an Antibody and is intended for research purposes only. The product is available for immediate purchase. The TCF4 (C48H11), Monoclonal Antibody, Rabbit reacts with Chicken, Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's TCF4 (C48H11) can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunoprecipitation (IP). Western Blot: 1:1000 Immunoprecipitation: 1:50 Dilutions listed only as a recommendation. Optimal dilution should be determined by investigator. Western Blot: Sample Preparation 1. Treat cells by adding fresh media containing regulator for desired time. 2. Aspirate media from cultures; wash cells with 1X PBS; aspirate. 3. Lyse cells by adding 1X SDS sample buffer (100 ul per well of 6-well plate or 500 ul per plate of 10 cmplate). Immediately scrape cells from plate and transfer the extract to a microcentrifuge tube. Keep on ice. 4. Sonicate for 10–15 seconds to shear DNA and reduce sample viscosity. 5. Heat a 20 ul sample to 95–100°C for 5 minutes; cool on ice. 6. Microcentrifuge for 5 minutes. 7. Load 20 ul onto SDS-PAGE gel (10 cm x 10 cm). 8. Electrotransfer to nitrocellulose (or PVDF) membrane. Membrane Blocking and Antibody Incubations Note: Volumes for 10 cm x 10 cm (100 cm2) membrane; for different sized membranes, adjust vol. accordingly. 1. (Optional) After transfer, wash nitrocellulose membrane with 25 ml TBS for 5 minutes at room temperature. 2. Incubate membrane in 25 ml of blocking buffer for 1 hour at room temperature. 3. Wash three times for 5 minutes each with 15 ml of TBS/T. 4. Incubate membrane and primary antibody (at the appropriate dilution) in 10 ml primary antibody dilutionbuffer with gentle agitation overnight at 4°C. 5. Wash three times for 5 minutes each with 15 ml of TBS/T. 6. Incubate membrane with HRP-conjugated secondary antibody (1:2000) in 10 ml of blocking buffer withgentle agitation for 1 hour at room temperature. 7. Wash three times for 5 minutes each with 15 ml of TBS/T. 8. Process membranes using enhanced chemiluminescence. Immunoprecipitation: Preparing Cell Lysates 1. Aspirate media. Treat cells by adding fresh media containing regulator fordesired time. 2. To harvest cells under nondenaturing conditions, remove media and rinse cells once with ice-cold PBS. 3. Remove PBS and add 0.5 ml ice-cold 1X cell lysis buffer to each plate (10 cm)and incubate the plates on ice for 5 minutes. 4. Scrape cells off the plates and transfer to microcentrifuge tubes. Keep on ice. 5. Sonicate samples on ice three times for 5 seconds each. 6.Microcentrifuge for 10 minutes at 14,000 X g, 4°C, and transfer the supernatantto a new tube. If necessary, lysate can be stored at –80°C. Immunoprecipitation Optional: It may be necessary to perform a lysate pre-cleaning step to reduce non-specificbinding to the Protein A/G agarose beads (See section below). 1. Take 200 ul cell lysate and add primary antibody. Incubate with gentle rockingovernight at 4°C. 2. Add either protein An or G agarose beads (20 ul of 50% bead slurry). Incubate with gentle rocking for 1–3 hours at 4°C. 3. Microcentrifuge for 30 seconds at 4°C. Wash pellet five times with 500 ul of 1Xcell lysis buffer. Keep on ice during washes. 4. Resuspend the pellet with 20 ul 3X SDS sample buffer. Vortex, then microcentrifugefor 30 seconds. 5. Heat the sample to 95–100°C for 2–5 minutes and microcentrifuge for 1 minuteat 14,000 X g. 6. Load the sample (15–30 ul) on SDS-PAGE gel (12–15%). 7. Analyze sample by Western blotting (see Western Immunoblotting Protocol). Cell Lysate Pre-Cleaning (Optional) 1. Take 200 ul cell lysate and add to either Protein An or G agarose beads (20 ul of50% bead slurry). 2. Incubate at 4°C for 30 – 60 minutes. 3. Spin for 10 minutes at 4°C. Transfer the supernatant to a fresh tube. 4. Proceed to step 1 of Immunoprecipitation. Researchers should empirically determine the suitability of the TCF4 tcf4 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 "TCF4 (C48H11), 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.Item has been added to Shopping Cart
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