Invitrogen

POLR1D Polyclonal Antibody

Advanced Verification

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

1 Published Figure

1 Reference

View all (8) POLR1D antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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POLR1D Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of POLR1D was performed using 70% confluent log phase A549 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 POLR1D Polyclonal Antibody (Product # PA5-30575) at 1:200 dilution 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). 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.

POLR1D Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of paraffin-embedded HBL435 xenograft, using POLR1D (Product # PA5-30575) antibody at 1:500 dilution. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

POLR1D Antibody in Western Blot (WB) — Knockdown of POLR1D was achieved by transfecting HeLa cells with POLR1D specific siRNAs (Silencer® select Product # s234268). Western blot analysis (Fig. a) was performed using whole cell extracts from the POLR1D knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with POLR1D Polyclonal Antibody (Product # PA5-30575, 1:3000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/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 POLR1D.

POLR1D Antibody in ChIP Assay (ChIP) — Chromatin immunoprecipitation analysis of POLR1D was performed using cross-linked chromatin from 1x10^6 HCT116 colon carcinoma cells treated with serum for 0 and 30 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of a POLR1D polyclonal antibody (Product # PA5-30575). Chromatin aliquots from ~1x10^5 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify exon-1 of the RNA5-8S5 gene or -20 kb upstream of the LAMC1 gene. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representations of the RNA5-8S5 and LAMC1 loci are shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by RNA5-8S5 and LAMC1 primers are represented by black bars. Data courtesy of the Innovators Program.

POLR1D Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with POLR1D siRNA and reduction of signal was observed in Western Blot using POLR1D Polyclonal Antibody (Product # PA5-30575). {KD}

Product Details

PA5-30575

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	View 1 publication 1 publication
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	1:200	-
ChIP assay (ChIP)	Assay-Dependent	-

Product Specifications
Species Reactivity	Human
Published species	Human
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Full length human POLR1D (AAH00889) Recombinant protein.
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	0.1M tris glycine, 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_2548049

Product Specific Information

Recommended positive controls: A549, HeLa, HepG2, HCT116.

Predicted reactivity: Mouse (90%), Rhesus Monkey (99%), Bovine (94%).

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

Target Information

The protein encoded by this gene is a component of the RNA polymerase I and RNA polymerase III complexes, which function in the synthesis of ribosomal RNA precursors and small RNAs, respectively. Mutations in this gene are a cause of Treacher Collins syndrome (TCS), a craniofacial development disorder. Alternative splicing results in multiple transcript variants.

⚠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.

Product References

Interaction between the BAG1S isoform and HSP70 mediates the stability of anti-apoptotic proteins and the survival of osteosarcoma cells expressing oncogenic MYC.

BMC cancer

Gennaro VJ,Wedegaertner H,McMahon SB

PA5-30575 was used in Western Blotting to identify a set of specific pro-survival clients controlled by the HSP70/BAG1S chaperone complex.

Fri Mar 22 00:00:00 EDT 2019

Interaction between the BAG1S isoform and HSP70 mediates the stability of anti-apoptotic proteins and the survival of osteosarcoma cells expressing oncogenic MYC.

BMC cancer

Gennaro VJ,Wedegaertner H,McMahon SB

PA5-30575 was used in Western Blotting to identify a set of specific pro-survival clients controlled by the HSP70/BAG1S chaperone complex.

Fri Mar 22 00:00:00 EDT 2019

Bioinformatics

Protein Aliases: AC19; DNA-directed RNA polymerase I subunit D; DNA-directed RNA polymerases I and III subunit RPAC2; hRPA19; polymerase (RNA) I polypeptide D; polymerase (RNA) I polypeptide D, 16kDa; polymerase (RNA) I subunit D; Protein POLR1D, isoform 2; RNA polymerase I 16 kDa subunit; RNA polymerases I and III subunit AC2; RPA16; RPC16

Gene Aliases: AC19; POLR1C; POLR1D; RPA16; RPA9; RPAC2; RPC16; RPO1-3; TCS2

UniProt ID: (Human) P0DPB6

Entrez Gene ID: (Human) 51082

Molecular Function: DNA-directed RNA polymerase RNA metabolism protein nucleic acid metabolism protein

Function(s)

transcription initiation from RNA polymerase I promoter transcription elongation from RNA polymerase I promoter termination of RNA polymerase I transcription transcription from RNA polymerase III promoter positive regulation of type I interferon production positive regulation of gene expression, epigenetic

Process(es)

RNA polymerase I activity RNA polymerase III activity DNA binding protein binding protein dimerization activity

POLR1D Polyclonal Antibody

Product Details

PA5-30575

Western Blot (WB)

Immunohistochemistry (Paraffin) (IHC (P))

Immunocytochemistry (ICC/IF)

ChIP assay (ChIP)

Species Reactivity

Published species

Host/Isotype

Class

Type

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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