Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Flow cytometry [1]
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Validation data
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- Product number
- 40-2400 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Arfaptin 2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.25 mg/mL
- Storage
- -20°C
Submitted references Structural basis for membrane binding specificity of the Bin/Amphiphysin/Rvs (BAR) domain of Arfaptin-2 determined by Arl1 GTPase.
Arfaptins are localized to the trans-Golgi by interaction with Arl1, but not Arfs.
Nakamura K, Man Z, Xie Y, Hanai A, Makyio H, Kawasaki M, Kato R, Shin HW, Nakayama K, Wakatsuki S
The Journal of biological chemistry 2012 Jul 20;287(30):25478-89
The Journal of biological chemistry 2012 Jul 20;287(30):25478-89
Arfaptins are localized to the trans-Golgi by interaction with Arl1, but not Arfs.
Man Z, Kondo Y, Koga H, Umino H, Nakayama K, Shin HW
The Journal of biological chemistry 2011 Apr 1;286(13):11569-78
The Journal of biological chemistry 2011 Apr 1;286(13):11569-78
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed using membrane enriched extracts (30 µg) of Mouse Heart (Lane 1), nuclear enriched extract of MDA-MB-231 (Lane 2), A-431 (Lane 3), Hep G2 (Lane 4), HEK 293 (Lane 5), and membrane enriched HT-29 (Lane 6), Rat Brain (Lane 7) and nuclear enriched extract of HeLa (Lane 8).The blots were probed with Anti-Arfaptin 2 Rabbit Polyclonal Antibody (Product # 40-2400, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4µg/mL 1:2500 dilution). A ~34 kDa band corresponding to Arfaptin 2 was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed using membrane enriched extracts (30 µg) of Mouse Heart (Lane 1), nuclear enriched extract of MDA-MB-231 (Lane 2), A-431 (Lane 3), Hep G2 (Lane 4), HEK 293 (Lane 5), and membrane enriched HT-29 (Lane 6), Rat Brain (Lane 7) and nuclear enriched extract of HeLa (Lane 8).The blots were probed with Anti-Arfaptin 2 Rabbit Polyclonal Antibody (Product # 40-2400, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4µg/mL 1:2500 dilution). A ~34 kDa band corresponding to Arfaptin 2 was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Arfaptin 2 was performed using 70% confluent log phase MDA-MB-231 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Arfaptin 2 Rabbit Polyclonal Antibody (Product # 40-2400) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of Arfaptin 2 was performed using MCF7 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Arfaptin 2 Rabbit Polyclonal Antibody (Product # 40-2400, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10, 000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.