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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- PA5-116079 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TSC1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Antibody detects endogenous levels of total TSC1.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TSC1 in HeLa cells lysate (left lane: treated with blocking peptide). Samples were incubated with polyclonal antibody (Product # PA5-116079).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TSC1 in various samples (Lane 1: Mouse brain lysates; Lane 2: Mouse heart lysates; Lane 3: Mouse brain lysates; Lane 4: Mouse brain lysates treated with blocking peptide ). Samples were incubated with polyclonal antibody (Product # PA5-116079).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot of TSC1 was performed by loading 100µg of WT (lane 1) and TSC1 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. TSC1 was detected at approximately 130 kDa (designated by the black arrow) using a TSC1 polyclonal antibody (Product # PA5-116079) 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 # 65-6120) 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
- Immunocytochemistry analysis of TSC1 in HeLa cells. Samples were treated with PFA, permeabilized in 0.1% Triton X-100, blocked in 10% serum (45 min at 25°C), and incubated with polyclonal antibody (Product # PA5-116079) at a dilution of 1:200 (1 hr at 37°C). Secondary staining was applied with mouse anti-beta tubulin; AlexaFluor 594 conjugated goat anti-rabbit IgG (Red); AlexaFluor 88 conjugated goat anti-mouse IgG (Green) and DAPI (blue) using a dilution of 1:200 (1 hr at 37°C).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of TSC1 was performed on HAP1 cell lysates. Antibody-bead conjugates were prepared by adding 2 µg of TSC1 polyclonal antibody (Product # PA5-116079) with 30 µL of protein A-Sepharose beads and rocked overnight at 4°C. 1 mg 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 the same antibody. 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.
