PA1-072

antibody from Invitrogen Antibodies

Targeting: APBA3 mint3, X11L2

Antibody data

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Validation data

Product number: PA1-072 - Provider product page
Provider: Invitrogen Antibodies
Product name: MINT3 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA1-072 detects munc-18 interacting protein 3 (Mint3) from mouse cells and rat brain samples. PA1-072 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~86 kDa protein representing Mint3 from AtT20 cell extract and rat brain samples. The PA1-072 immunogen is a synthetic peptide corresponding to residues M(1) E F L P E P Q H P P G P P T M D L E(19) of rat Mint3. The PA1-072 immunizing peptide (Cat. # PEP-135) is available for use in neutralization and control experiments.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

APBA3 protein structure - PA1-072 shown in red.

Wu Q, Dai Q, Jiang L, Wang Y, Yang T, Miao J, Wang J, Han Y
Molecular medicine reports 2017 Aug;16(2):1578-1583

Sullivan SE, Dillon GM, Sullivan JM, Ho A
The Journal of biological chemistry 2014 May 30;289(22):15374-83

Chaufty J, Sullivan SE, Ho A
The Journal of neuroscience : the official journal of the Society for Neuroscience 2012 Jul 11;32(28):9613-25

Biederer T, Cao X, Südhof TC, Liu X
The Journal of neuroscience : the official journal of the Society for Neuroscience 2002 Sep 1;22(17):7340-51

Biederer T, Cao X, Südhof TC, Liu X
The Journal of neuroscience : the official journal of the Society for Neuroscience 2002 Sep 1;22(17):7340-51

Okamoto M, Nakajima Y, Matsuyama T, Sugita M
Neuroscience 2001;104(3):653-65

Nakajima Y, Okamoto M, Nishimura H, Obata K, Kitano H, Sugita M, Matsuyama T
Brain research. Molecular brain research 2001 Aug 15;92(1-2):27-42

Nakajima Y, Okamoto M, Nishimura H, Obata K, Kitano H, Sugita M, Matsuyama T
Brain research. Molecular brain research 2001 Aug 15;92(1-2):27-42

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot detection of Mint3 from AtT20 cell extract using Product # PA1-072.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot detection of Mint3 from AtT20 cell extract using Product # PA1-072.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of U-2 OS (Lane 1), HCT 116 (Lane 2) and DU 145 (Lane 3). The blots were probed with Anti-Mint3 Rabbit Polyclonal Antibody (Product # PA1-072, 2 µg/mL) 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 62 kDa band corresponding to Mint3 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with 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 Mint3 was performed using 90% confluent log phase Neuro-2a 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 Mint3 Rabbit Polyclonal Antibody (Product # PA1-072) 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) at 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 membranous 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 Mint3 was done on Neuro-2a 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 Mint3 Rabbit Polyclonal Antibody (PA1-072, 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 (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..

Supportive validation