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Western blot
ImmunohistochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunohistochemistry [4]
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- Product number
- LS-C756394 - Provider product page
- Provider
- LSBio
- Product name
- ATF4 Antibody LS-C756394
- Antibody type
- Polyclonal
- Description
- Immunogen affinity purified
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Storage
- After reconstitution, may be stored at 4°C for 1 month. For long-term storage and to avoid freeze-thaw cycles, aliquot and store at -20°C.
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Supportive validation
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- Western blot analysis of ATF4 using anti-ATF4 antibody. 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 placenta tissue lysates, Lane 3: human HepG2 whole cell lysates,Lane 4: human MDA-MB-231 whole cell lysates,Lane 5: human SW620 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 rabbit anti-ATF4 antigen affinity purified polyclonal antibody at 0.5 ?g/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 1:10000 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 ATF4 at approximately 49KD. The expected band size for ATF4 is at 39KD.
Supportive validation
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- IHC analysis of ATF4 using anti-ATF4 antibody. ATF4 was detected in paraffin-embedded section of human prostatic cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2?g/ml rabbit anti-ATF4 Antibody 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) with DAB as the chromogen.
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- IHC analysis of ATF4 using anti-ATF4 antibody. ATF4 was detected in paraffin-embedded section of human ovary cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2?g/ml rabbit anti-ATF4 Antibody 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) with DAB as the chromogen.
- Submitted by
- LSBio (provider)
- Main image
- Experimental details
- IHC analysis of ATF4 using anti-ATF4 antibody. ATF4 was detected in paraffin-embedded section of human ovary cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2?g/ml rabbit anti-ATF4 Antibody 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) with DAB as the chromogen.
- Submitted by
- LSBio (provider)
- Main image
- Experimental details
- IHC analysis of ATF4 using anti-ATF4 antibody. ATF4 was detected in paraffin-embedded section of human prostatic cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2?g/ml rabbit anti-ATF4 Antibody 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) with DAB as the chromogen.
