Invitrogen

PABP Polyclonal Antibody

Advanced Verification

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

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Datasheet

Protocols

Questions & Answers

Tech Support

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PABP Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of PABP was performed using 70% confluent log phase HCT 116 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PABP Rabbit Polyclonal Antibody(Product # PA5-29883) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight 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 ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). 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 represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

PABP Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — PABP Polyclonal Antibody detects PABP protein at cytosol on mouse middle brain by immunohistochemical analysis. Sample: Paraffin-embedded mouse middle brain. PABP Polyclonal Antibody (Product # PA5-29883) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PABP Antibody in Western Blot (WB) — Knockdown of PABP was achieved by transfecting HCT 116 cells with PABP specific siRNAs (Silencer® select Product # s25666, s25664). Western blot analysis (Fig. a) was performed using whole cell extracts from the PABP knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PABP Polyclonal Antibody (Product # PA5-29883, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 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 PABP.

PABP Antibody in Immunoprecipitation (IP) — Detection of binding of endogenous PABP protein to specific RNA using Anti-PABP Antibody: RNA Immunoprecipitation (RIP) was performed using Anti-PABP Polyclonal Antibody (Product # PA5-29883, 4 µg) on clarified whole cell lysate from 2 million Hep G2 cells. Normal Rabbit IgG was used as a negative IP control. Immunoprecipitated RNA was purified by RiboPure™ RNA Purification Kit (Product # AM1924) and analyzed by RT-PCR using the Power SYBR® Green RNA-to-CT™ 1-Step Kit (Product # 4389986) with primer pairs over ACTB mRNA, GAPDH mRNA (positive) and U1 snRNA, 18S rRNA (negative). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

PABP Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HCT 116 cells were transfected with PABP siRNA and reduction of signal was observed in Western Blot using PABP Polyclonal Antibody (Product # PA5-29883). {KD}

Product Details

PA5-29883

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	-

Product Specifications
Species Reactivity	Rat, Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant fragment corresponding to a region within amino acids 356 and 630 of Human PABP View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 20% glycerol
Contains	0.01% thimerosal
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2547357

Product Specific Information

Recommended positive controls: 293T, A431, HeLa, HepG2, A549, H1299, Neuro2A, C8D30, NIH-3T3, Raw264.7, C2C12, PC-12, Rat2.

Predicted reactivity: Mouse (99%), Rat (98%), Xenopus laevis (91%), Chicken (95%), Rhesus Monkey (100%), Bovine (100%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

The poly(A)-binding protein (PABP), which is found complexed to the 3-prime poly(A) tail of eukaryotic mRNA, is required for poly(A) shortening and translation initiation. In humans, the PABPs comprise a small nuclear isoform and a conserved gene family that displays at least 3 functional proteins: PABP1 (PABPC1), inducible PABP (iPABP, or PABPC4), and PABP3 (PABPC3). In addition, there are at least 4 pseudogenes, PABPCP1 to PABPCP4.

⚠WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause birth defects or other reproductive harm. For more information go to www.P65Warnings.ca.gov.

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

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Bioinformatics

Protein Aliases: PABP 1; PABP-1; poly A binding protein 1; poly(A) binding protein, cytoplasmic 1; poly(A) binding protein, cytoplasmic 2; poly(A)-binding protein 1; Polyadenylate-binding protein 1

Gene Aliases: ePAB; PAB1; PABP; PABP1; PABPC1; PABPC2; PabpI; PABPL1

UniProt ID: (Human) P11940, (Rat) Q9EPH8, (Mouse) P29341

Entrez Gene ID: (Human) 26986, (Rat) 171350, (Mouse) 18458

Function(s)

nuclear-transcribed mRNA catabolic process, nonsense-mediated decay mRNA processing RNA splicing gene silencing by RNA positive regulation of nuclear-transcribed mRNA poly(A) tail shortening positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay negative regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay nuclear-transcribed mRNA poly(A) tail shortening mRNA splicing, via spliceosome mRNA polyadenylation translational initiation regulation of mRNA stability positive regulation of viral genome replication positive regulation of translation mRNA stabilization

Process(es)

nucleotide binding protein binding protein C-terminus binding poly(A) binding poly(U) RNA binding translation activator activity poly(A) RNA binding mRNA binding nucleic acid binding RNA binding

PABP Polyclonal Antibody

Product Details

PA5-29883

Western Blot (WB)

Immunohistochemistry (Paraffin) (IHC (P))

Immunocytochemistry (ICC/IF)

Species Reactivity

Host/Isotype

Class

Type

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

References (0)

Bioinformatics

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