Invitrogen

Rab4 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

4 Published Figures

7 References

View all (9) Rab4 antibodies

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Datasheet

Protocols

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Rab4 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of RAB4A 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 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with RAB4A Rabbit polyclonal Antibody (Product # PA3-912) at 1:250 dilution 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 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.

Rab4 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on cancer biopsies of deparaffinized human Breast carcinoma tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a rabbit polyclonal antibody recognizing RAB4 (Product # PA3-912) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Rab4 Antibody in Western Blot (WB) — Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), NIH/3T3 (Lane 2), COS-7 (Lane 3), U-87 MG (Lane 4), Jurkat (Lane 5), K-562 (Lane 6), PC-12 (Lane 7) and A549 (Lane 8). The blot was probed with Rabbit Anti-RAB4A Polyclonal Antibody (Product # PA3-912, 1:500) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A 24 kDa band corresponding to RAB4A 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).

Rab4 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Fig. 5 FERMT2 silencing inhibits APP degradation and promotes APP recycling at the plasma membrane. a HEK293-APP 695WT cells transiently transfected with siFERMT2 or non-targeting siRNA were treated with bafilomycin A1 (BafA1, 50 nM) for the indicated times. Cell extracts were then analyzed by WB. Densitometric analyses and mature APP levels for three independent experiments are shown. Graphs indicate the mean +- SD. * p < 0.05, non-parametric test. b The time course of APP endocytosis and degradation was indirectly visualized by internalization of 6E10 antibody. Cells were incubated with 6E10 antibody at 4 degC for 1 h. The temperature was then shifted to 37 degC, and cells were processed for immunofluorescence at the indicated times. Scale bar 10 um. c Relative fluorescence intensity from 6E10 staining, showing the time course of APP degradation. d A zoom-in (the square in b ) for the indicated times (0 and 20 min at 37 degC). Co-staining with anti-Rab4 antibody was used to visualize the APP within Rab4-positive endosomes involving in recycling. e Co-localization of 6E10 staining with Rab4 staining, as a guide to the APP level within recycling endosomes at the indicated times. f Cells transiently transfected with anti-FERMT2 in the presence or absence of siRab4. Extracts were analyzed by WB using anti-APP C-terminal, anti-FERMT2, anti-Rab4 or anti-actin antibodies

Rab4 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with RAB4A specific validated siRNA (Silencer® select Product # s11675) and decrease in signal intensity was observed in western blot application using Anti-RAB4A Rabbit Polyclonal Antibody (Product # PA3-912). {KD}

Product Details

PA3-912

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500	View 3 publications 3 publications
Immunohistochemistry (IHC)	1:200	-
Immunocytochemistry (ICC/IF)	1:250	View 4 publications 4 publications

Product Specifications
Species Reactivity	Dog, Human, Mouse, Non-human primate, Rat
Published species	Human
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Purified, recombinant human Rab4 protein.
Conjugate	Unconjugated
Form	Liquid
Concentration	Conc. Not Determined
Storage buffer	whole serum
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2269382

Product Specific Information

PA3-912 detects Rab4 from human and mouse samples. This antibody does not detect other Rab isoforms (Rab1, Rab3, Rab5, or Rab6).

PA3-912 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects a ~24 kDa band representing Rab4 from transfected HeLa cells.

The PA3-912 immunogen is purified recombinant human Rab4 protein.

Target Information

Protein transport. Probably involved in vesicular traffic.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

RAB5A expression is a predictive biomarker for trastuzumab emtansine in breast cancer.

Nature communications

Engebraaten O,Yau C,Berg K,Borgen E,Garred Ø,Berstad MEB,Fremstedal ASV,DeMichele A,Veer LV',Esserman L,Weyergang A

Published figure using Rab4 polyclonal antibody (Product # PA3-912) in Western Blot

Fri Nov 05 00:00:00 EDT 2021

Nuclear and Cytoplasmatic Quantification of Unconjugated, Label-Free Locked Nucleic Acid Oligonucleotides.

Nucleic acid therapeutics

Pendergraff H,Schmidt S,Vikeså J,Weile C,Øverup C,W Lindholm M,Koch T

PA3-912 was used in Western Blot to show compound variability (diversity) by which 247,000-693,000 LNAs/nuclei results in similar target reduction for different compounds.

Sat Feb 01 00:00:00 EST 2020

WDFY2 restrains matrix metalloproteinase secretion and cell invasion by controlling VAMP3-dependent recycling.

Nature communications

Sneeggen M,Pedersen NM,Campsteijn C,Haugsten EM,Stenmark H,Schink KO

Published figure using Rab4 polyclonal antibody (Product # PA3-912) in Immunocytochemistry

Fri Jun 28 00:00:00 EDT 2019

Endosomal trafficking defects in patient cells with KIAA1109 biallelic variants.

Genes & diseases

Kane MS,Diamonstein CJ,Hauser N,Deeken JF,Niederhuber JE,Vilboux T

PA3-912 was used in Immunocytochemistry-immunoflourescence to study the mechanism by which KIAA1109 loss of function causes this spectrum of congenital neurological malformation disorders.

Fri Mar 01 00:00:00 EST 2019

Genome-wide, high-content siRNA screening identifies the Alzheimer's genetic risk factor FERMT2 as a major modulator of APP metabolism.

Acta neuropathologica

Chapuis J,Flaig A,Grenier-Boley B,Eysert F,Pottiez V,Deloison G,Vandeputte A,Ayral AM,Mendes T,Desai S,Goate AM,Kauwe JSK,Leroux F,Herledan A,Demiautte F,Bauer C,Checler F,Petersen RC,Blennow K,Zetterberg H,Minthon L,Van Deerlin VM,Lee VM,Shaw LM,Trojanowski JQ,Albert M,Moghekar A,O'Brien R,Peskind ER,Malmanche N,Schellenberg GD,Dourlen P,Song OR,Cruchaga C,Amouyel P,Deprez B,Brodin P,Lambert JC

Published figure using Rab4 polyclonal antibody (Product # PA3-912) in Immunofluorescence

Thu Jun 01 00:00:00 EDT 2017

Genome-wide, high-content siRNA screening identifies the Alzheimer's genetic risk factor FERMT2 as a major modulator of APP metabolism.

Acta neuropathologica

Published figure using Rab4 polyclonal antibody (Product # PA3-912) in Immunofluorescence

Thu Jun 01 00:00:00 EDT 2017

Dynamic internalization and recycling of a metal ion transporter: Cu homeostasis and CTR1, the human Cu⁺ uptake system.

Journal of cell science

Clifford RJ,Maryon EB,Kaplan JH

PA3-912 was used in immunocytochemistry study how importation of copper into cells is regulated

Fri Apr 15 00:00:00 EDT 2016

Bioinformatics

Protein Aliases: HTLV-1 related endogenous sequence; Ras-related protein Rab-4A; Ras-related small GTP binding protein 4

Gene Aliases: AI848268; HRES-1; HRES-1/RAB4; HRES1; RAB4; RAB4A

UniProt ID: (Human) P20338, (Rat) P05714, (Mouse) P56371

Entrez Gene ID: (Human) 5867, (Dog) 479210, (Rat) 25532, (Mouse) 19341

Function(s)

protein transport antigen processing and presentation regulation of endocytosis Rab protein signal transduction positive regulation of ATPase activity intracellular protein transport small GTPase mediated signal transduction metabolic process transport

Process(es)

ATPase activator activity GTPase activity protein binding GTP binding protein transporter activity GDP binding syntaxin binding ionotropic glutamate receptor binding ATPase binding nucleotide binding

Rab4 Polyclonal Antibody

Product Details

PA3-912

Western Blot (WB)

Immunohistochemistry (IHC)

Immunocytochemistry (ICC/IF)

Species Reactivity

Published species

Host/Isotype

Class

Type

Immunogen

Conjugate

Form

Concentration

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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