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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Other assay [1]
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- Product number
- PA5-53581 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Spastin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: KRKDPLTHTS NSLPRSKTVM KTGSAGLSGH HRAPSYSGLS MVSGVKQGSG PAPTTHKGTP KTNRTNKPST PTTATRKKKD LKNFRNVDSN LANLIMNEIV DNGTAVKFDD IAGQDLAKQA LQEIVILPSL RPELFTGL
- Concentration
- 0.10 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Spastin in Lane 1: Marker (kDa) 230, 130, 95, 72, 56, 36, 28, 17, 11; Lane 2: Human cell line RT-4. Samples were probed using a Spastin Polyclonal Antibody (Product # PA5-53581).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Spastin in Lane 1: NIH-3T3 cell lysate (Mouse embryonic fibroblast cells); Lane 2: NBT-II cell lysate (Rat Wistar bladder tumour cells). Samples were probed using a Spastin Polyclonal Antibody (Product # PA5-53581).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot of SPAST was performed by loading 80 µg of WT (lanes 1/3) and SPAST CRISPR KO (lanes 2/4) HEK293T or HAP1 cell lysates in RIPA buffer onto a 4-15% 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. SPAST was detected at approximately 67 kDa (designated by the black arrow) using a SPAST polyclonal antibody (Product # PA5-53581) 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
- Immunofluorescent staining of Spastin in human cell line U-251 MG using a Spastin Polyclonal Antibody (Product # PA5-53581) shows localization to nucleoplasm and cytosol.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical staining of Spastin in human kidney tissue shows moderate positivity in cytoplasm and membranes of tubular cells. Samples were probed using a Spastin Polyclonal Antibody (Product # PA5-53581).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of SPAST was performed on HAP1 cell lysates. Antibody-bead conjugates were prepared by adding 1 µg of SPAST polyclonal antibody (Product # PA5-53581) 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 a different SPAST monoclonal 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.
