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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- MA5-32675 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SFRP1 Recombinant Rabbit Monoclonal Antibody (JA11-68)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JA11-68
- 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
- Immunofluorescence analysis of SFRP1 was performed using 70% confluent log phase MCF 10A and MCF7 cells untreated as well as PTI treated. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with SFRP1 Recombinant Rabbit Monoclonal Antibody (JA11-68) (Product # MA5-32675) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790, 1:2000 dilution), for 45 minutes at room temperature (Panel a, e, i, m: Green). Nuclei (Panel b, f, j, n: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c, g, k, o: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel h represents the merged image showing cytoplasmic as well as Golgi like localization. Panel (a-d) represent MCF 10A cells. Panel (e-h) represents MCF 10A cells treated with PTI (1X for 4hrs). Panel (i-l) represents MCF7 cells. Panel (m-p) represents MCF7 treated with PTI The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- A549 cells were washed 3x with PBS and starved for ~18 hrs. Immunoprecipitation of SFRP1 was performed on A549 concentrated cell culture media, concentrated by centrifuging for 10 min at 500 x g to eliminate cells and larger contaminants, then for 10 min at 4,500 x g to eliminate smaller contaminants. Culture media were concentrated using a centrifugal filter with a membrane NMWL of 10 kDa at 4,000 x g for 10 min. Antibody-bead conjugates were prepared by adding 1 µg of SFRP1 recombinant monoclonal antibody (Product # MA5-32675) 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 SFRP1 recombinant monoclonal antibody (Product # MA5-38193). 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.
