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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Flow cytometry [1]
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Validation data
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- Product number
- MA1-172 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- RAB27A Monoclonal Antibody (4D3F11)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 4D3F11
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of RAB27A was performed by loading 20 µg of SK-MEL-28 and SH-SY5Y whole cell lysates and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a Novex 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BOX ). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. RAB27A was detected at ~25 kDa using a RAB27A monoclonal antibody (Product # MA1-172) at a dilution of 1 µg/mL in 5% Milk in TBST overnight at 4°C on a rocking platform, followed by a Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A28177) at a dilution of 1:1,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal Pico substrate (Product # 34078) and the myECL Imager (Product # 62236).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of RAB27A was done on HeLa cells. Cells were fixed, permeabilized and stained with a RAB27A mouse monoclonal antibody (Product # MA1-172, green histogram) or an isotype control (Product # MA5-14447, black histogram) at a dilution of 10 µg/mL. After incubation for 1 hour on ice, the cells were labeled with Goat anti-Mouse IgG Secondary Antibody, DyLight 650 conjugate (Product # 84545) at a dilution of 1:50 for 1 hour on ice. A representative 10,000 cells were acquired and analyzed for each sample.
