PA1-770

antibody from Invitrogen Antibodies

Targeting: RAB3A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-770

Provider

Invitrogen Antibodies

Product name

RAB3A Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA1-770 detects rab 3A from canine, hamster, human, mouse and rat tissues and cells. PA1-770 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot, this antibody detects an ~21 kDa protein representing rab 3A from Hepa1 cell lysate. PA1-770 immunogen is a synthetic peptide corresponding to residues M(1) A S A T D S R Y G Q K E S S D Q N(18) C of human RAB3A. This sequence is completely conserved between human, mouse, and rat rab 3A. PA1-770 immunizing peptide (Cat. # PEP-098) is available for use in neutralization and control experiments. Recombinant human rab 3A (Cat # RP-770) protein can be purchased for control experiments in Western blot applications.

Reactivity

Human, Mouse, Rat, Canine, Hamster

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Secreted key regulators (Fgf1, Bmp4, Gdf3) are expressed by PAC1-immunopositive retinal ganglion cells in the postnatal rat retina.

Dénes V, Kovacs K, Lukáts Á, Mester A, Berta G, Szabó A, Gabriel R
European journal of histochemistry : EJH 2022 Apr 27;66(2)

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Systematic proteomic analysis of LRRK2-mediated Rab GTPase phosphorylation establishes a connection to ciliogenesis.

Steger M, Diez F, Dhekne HS, Lis P, Nirujogi RS, Karayel O, Tonelli F, Martinez TN, Lorentzen E, Pfeffer SR, Alessi DR, Mann M
eLife 2017 Nov 10;6

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-RAB3A Polyclonal Antibody (Product # PA1-770) and a 27 kDa band corresponding to RAB3A was observed along with two uncharacterized band (*) at ~40 kD and 60 kDa across tissues tested. Tissue extract (30 µg lysate) of Mouse Brain (Lane 1), Mouse Kidney (Lane 2), Mouse Liver (Lane 3), Rat Brain (Lane 4), Rat Kidney (Lane 5), Rat Liver (Lane 6), were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (2 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of RAB3A was done on 70% confluent log phase HeLa 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 RAB3A Rabbit Polyclonal Antibody (Product # PA1-770) at 2 µg/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 is a merged image showing Cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of RAB3A was done on HeLa 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 RAB3A Rabbit Polyclonal Antibody (PA1770, red histogram) or with rabbit isotype control (yellow histogram) at 3-5 ug/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.