Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
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- Product number
- PA5-77880 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Blooms Syndrome Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Positive Control: 293T, A431, Jurkat Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of BLM in A) 293T and B) A431 cells using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with BLM polyclonal antibody (Product # PA5-77880) using a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Blooms Syndrome Polyclonal Antibody (Product # PA5-77880). Whole cell extract (30 µg) was separated by 5% SDS-PAGE, and the membrane was blotted with Blooms Syndrome Polyclonal Antibody (Product # PA5-77880) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of Blooms Syndrome was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR815029_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Blooms Syndrome was performed by loading 30 µg of HeLa Cas9 (Lane 1) andHeLa Blooms Syndrome KO (Lane 2) modified whole cell extracts. The samples were electrophoresed using NuPAGE™ 3 to 8%, Tris-Acetate, 1.0 mm, Mini Protein Gel (Product # EC6695BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-Blooms Syndrome Polyclonal Antibody (Product # PA5-77880, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:5000 dilution) using the iBright™ FL 1500 (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Blooms Syndrome. Uncharacterized band was observed in all the samples at ~37 kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Blooms Syndrome Polyclonal Antibody detects BLM protein at cytoplasm and nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: BLM protein stained by Blooms Syndrome Polyclonal Antibody (Product # PA5-77880) diluted at 1:500. Red: phalloidin, a cytoskeleton marker, diluted at 1:200. Scale bar = 10 µm.