PA5-30234

antibody from Invitrogen Antibodies

Targeting: AGFG1 HRB, RAB, RIP

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-30234

Provider

Invitrogen Antibodies

Product name

AGFG1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: IMR32. Predicted reactivity: Mouse (98%), Rat (98%), Xenopus laevis (88%), Bovine (98%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

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.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using AGFG1 Polyclonal Antibody (Product # PA5-30234). Sample (30 µg of whole cell lysate). Lane A: IMR32 . 7.5% SDS PAGE. AGFG1 Polyclonal Antibody (Product # PA5-30234) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-AGFG1 Polyclonal Antibody (Product # PA5-30234) and a 60kDa band corresponding to AGFG1 was observed in IMR32, K562, A549 and Mouse Testis. An uncharacterized band (*), corresponding to ~90kDa were observed in the samples tested. Whole cell extracts (30 µg lysate) of IMR-32 (Lane 1), K-562 (Lane 2), A549 (Lane 3) and Mouse Testis (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 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

Immunofluorescent analysis of RIP/Rab in paraformaldehyde-fixed HeLa cells using a RIP/Rab polyclonal antibody (Product # PA5-30234) at a 1:500 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of AGFG1 was performed using 70% confluent log phase HeLa cells. 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 10 minutes at room temperature. The cells were labeled with AGFG1 Polyclonal Antibody (Product # PA5-30234) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with DAPI. F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using RIP/Rab (Product # PA5-30234) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.