PA5-30143

antibody from Invitrogen Antibodies

Targeting: RANGAP1 Fug1, KIAA1835, SD

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-30143

Provider

Invitrogen Antibodies

Product name

RANGAP1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: 293T, Neuro2A, C8D30, NIH-3T3, Raw264.7, C2C12, PC-12, Rat2. Predicted reactivity: Mouse (96%), Rat (95%), Xenopus laevis (86%), Chicken (90%), Bovine (96%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat

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 analysis of RanGAP1 using 30 µg of 293T lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a RanGAP1 polyclonal antibody (Product # PA5-30143) at a dilution of 1:5000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of RANGAP1 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a RANGAP1 Polyclonal Antibody (Product # PA5-30143) at a dilution of 1:2000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of RANGAP1 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a RANGAP1 Polyclonal Antibody (Product # PA5-30143) at a dilution of 1:2000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of RANGAP1 was performed by separating 30 µg of Whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a RANGAP1 Polyclonal Antibody (Product # PA5-30143) at a dilution of 1:5000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of RANGAP1 was achieved by transfecting HEK-293 with RANGAP1 specific siRNAs (Silencer® select Product # s11776, s11777). Western blot analysis (Fig. a) was performed using whole cell extracts from the RANGAP1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RANGAP1 Polyclonal Antibody (Product # PA5-30143, 1:5000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to RANGAP1..

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-RANGAP1 Polyclonal Antibody (Product # PA5-30143) and 63 kDa band corresponding to RANGAP1 was observed along with ~70, 80 kDa bands corresponding to SUMOylated form of RANGAP1 was observed across the cell lines tested. Whole cell Extracts of (30ug lysate) of K-562 (Lane 1), HEK-293 (Lane 2), HeLa (Lane 3), 3T3-L1 (Lane 4) , OVCAR-3 (Lane 5),Raji (Lane 6) and Mouse Testis (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1:5000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (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) (https://doi.org/10.1016/S0092-8674(00)81862-0)..