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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Other assay [1]
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- Product number
- AHO1422 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TSC2 Monoclonal Antibody (75R23)
- Antibody type
- Monoclonal
- Antigen
- Recombinant protein fragment
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 75R23
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Proteins from cell extracts of human HeLa cells (lane 1), mouse L929 cells (lane 2), and rat PC12 cells (lane 3) were resolved by SDS-PAGE and transferred to PVDF.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Proteins from cell extracts of human HeLa cells (lane 1), mouse L929 cells (lane 2), and rat PC12 cells (lane 3) were resolved by SDS-PAGE and transferred to PVDF.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot of TSC2 was performed by loading 100 µg of WT (lane 1) and TSC2 CRISPR KO (lane 2) HAP1 cell lysates in RIPA buffer onto a 3-12% gradient polyacrylamide gel. Proteins on the blots were visualized with Ponceau staining (below immunoblot). Proteins were transferred to nitrocellulose membrane and blocked in 5% milk for 1 hr. TSC2 was detected at approximately 200 kDa (designated by the black arrow) using a TSC2 monoclonal antibody (Product # AHO1422) at a dilution of 1:500 in 5% BSA in TBS with 0.1% Tween 20 (TBST) overnight at 4°C. The peroxidase-conjugated secondary antibody (Product # 62-6520) was diluted to 0.2 µg/mL in TBST with 5% milk for 1 hr. Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Product # 32106). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of TSC2 was performed on HAP1 cell lysates. Antibody-bead conjugates were prepared by adding 2 µg of TSC2 monoclonal antibody (Product # AHO1422) with 30 µL of protein A-Sepharose beads and rocked overnight at 4°C. 500 µg of lysate was incubated with an antibody-bead conjugate for 2 hours at 4°C. Following centrifugation and multiple washes, 10% starting material (SM), 10% unbound fraction (UB) and immunoprecipitated fraction (IP) were processed for immunoblot using another TSC2 antibody (Product # MA5-32190). Ponceau stained transfer of blot is shown. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.
