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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Chromatin Immunoprecipitation [1]
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- Product number
- 712019 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- BOB-1 Recombinant Polyclonal Antibody (2HCLC)
- Antibody type
- Polyclonal
- Antigen
- Other
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 2HCLC
- Vial size
- 100 µg
- Concentration
- 0.5 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 was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of Ramos (Lane 1), Raji (Lane 2), Daudi (Lane 3), Jurkat (Lane 4) and MOLT-4 (Lane 5). The blot was probed with Anti-POU2AF1 Recombinant Rabbit Polyclonal Antibody (Product # 712019, 1:2500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 35 kDa band corresponding to POU2AF1 was observed in relevant cell lines tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis,Raji cells were fixed and permiabilized detection of endogenous POU2AF1 using Anti-POU2AF1 Recombinant Rabbit Polyclonal Antibody (Product # 712019, 1:100) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Panel a-d) clearly demonstares predominant nuclear localization of POU2AF1 in Raji cells . Panel e-h) shows no specific staining in Jurkat cells which is POU2AF1 negative,demonstrating specificity. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Enrichment of endogenous POU2AF1 protein at specific gene loci using Anti-POU2AF1 Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-POU2AF1 Recombinant Rabbit Polyclonal Antibody (Product # 712019, 4 µg) on sheared chromatin from 2 million Ramos cells using MAGnify ChIP System (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over the promoters of TNFRSF17, IFIT3, BCL6, IFITM1 (active) and SAT2 satellite repeats (inactive). Data is presented as %IP (IP/Input) of the antibody signal versus the negative control IgG using the comparative CT method.
