PABP Antibody (10E10) - BSA Free

Western Blot: PABP Antibody (10E10) [NB120-6125] - Analysis of PABP expression in 1) A-431, 2) HeLa and 3) NIH-3T3 whole cell lysates.

Immunocytochemistry/Immunofluorescence: PABP Antibody (10E10) [NB120-6125] - PABP antibody was tested in HeLa cells at a 1:50 dilution using a Dylight 488 conjugated secondary antibody (Green). Actin (Red) and DNA (Blue) were counterstained using Phalloidin 568 and DAPI.

Flow Cytometry: PABP Antibody (10E10) [NB120-6125] - An intracellular stain was performed on A431 cells with PABP [10E10] Antibody NB120-6125AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Western Blot: PABP Antibody (10E10) [NB120-6125] - Cell line lysates were separated on SDS-PAGE and probed with 1 ug/mL Monoclonal Anti-PABP Clone: 10E10. The antibody was developed using Goat Anti-Mouse IgG-Peroxidase and a chemiluminescent substrate. (1) HEK293T, (2) HeLa, (3) G361 and (4) COS-7.

• Reactivity: Hu, Bv, Ca, Pm, Xp, Dr(-), Mu(-)
• Applications: WB, ELISA, Flow, ICC/IF, IP, MA, CyTOF-ready
• Clone: 10E10
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

PABP Antibody (10E10) - BSA Free Summary

• Immunogen: Recombinant human PABP protein. [Uniprot# P11940]
• Localization: Cytoplasmic. Shuttles between the cytoplasm and the nucleus.
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: PABPC1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • ELISA 1:100-1:2000
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:50-1:100
  • Immunoprecipitation 1:10-1:500
  • Microarray
  • Western Blot 1:1000
• Application Notes: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. This antibody is CyTOF ready.
• Theoretical MW: 69 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  A-431 Whole Cell Lysate
  HeLa Whole Cell Lysate

Reactivity Notes

Does not cross-react with Drosophilia melanogaster or Mouse. Not yet tested in other species.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for PABP Antibody (10E10) - BSA Free

• PAB1polyadenylate-binding protein 1
• PABP
• PABP-1
• PABP1poly(A) binding protein, cytoplasmic 2
• PABPC2
• PABPL1
• poly(A) binding protein, cytoplasmic 1
• Poly(A)-binding protein 1
• poly(A)-binding protein, cytoplasmic 2

Background

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. Grange et al. (1987) isolated a melanoma cell cDNA encoding human PABP. The predicted 633-amino acid protein contains 4 repeats of an approximately 80-amino acid unit in its N-terminal half. The authors found that this repeat region is highly conserved between human and yeast PABP and is sufficient for poly(A) binding. In vitro translation of the human PABP cDNA yielded a protein with an apparent molecular mass of 73 kD by SDS-PAGE. Northern blot analysis indicated that PABP is expressed as a 2.9-kb mRNA in human melanoma cells. Gorlach et al. (1994) noted that each of the 4 repeats of PABP is a ribonucleoprotein (RNP) consensus sequence RNA-binding domain. They determined that PABP has a pI of approximately 10.3 and is a very abundant, stable protein. Immunofluorescence studies of mammalian cells indicated that PABP is located exclusively in the cytoplasm. However, using both indirect immunofluorescence and tagging of PABP1 by fusion to the green fluorescent protein (GFP), Afonina et al. (1998) demonstrated that PABP1 shuttles between the nucleus and cytoplasm. PABP1 accumulated in the nucleus when transcription was inhibited, suggesting that active transcription is required for nuclear export of PABP1.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for PABP Antibody (NB120-6125)(9)

Publications using NB120-6125

• Kim B, Park Y, Hwang HJ et al. Single polysome analysis of mRNP Biochemical and biophysical research communications 2022-06-08 [PMID: 35716598]
• Genois MM, Gagné JP, Yasuhara T, et al. CARM1 regulates replication fork speed and stress response by stimulating PARP1 Molecular cell 2020-12-30 [PMID: 33412112]
• Chazal PE, Daguenet E, Wendling C et al. EJC core component MLN51 interacts with eIF3 and activates translation Proc Natl Acad Sci U S A 2013-03-25 [PMID: 23530232] (WB, Human)
• Sagliocco F, Laloo B, Cosson B et al. The ARE-associated factor AUF1 binds poly(A) in vitro in competition with PABP. Biochem J. 2006-12-01 [PMID: 16834569] (WB, Mouse)
• Larralde O, Smith RW, Wilkie GS et al. Direct stimulation of translation by the multifunctional herpesvirus ICP27 protein. J Virol. 2006-02-01 [PMID: 16415034] (WB, Human)
• Nelson PT, Hatzigeorgiou AG, Mourelatos Z. miRNP:mRNA association in polyribosomes in a human neuronal cell line. RNA. 2004-03-01 [PMID: 14970384] (WB, Human)
• Sladic RT, Lagnado CA, Bagley CJ, Goodall GJ. Human PABP binds AU-rich RNA via RNA-binding domains 3 and 4. Eur J Biochem. 2004-01-01 [PMID: 14717712] (IP, Human)
• Lee J, Bedford MT. PABP1 identified as an arginine methyltransferase substrate using high-density protein arrays. EMBO Rep. 2002-03-01 [PMID: 11850402] (IP, Human)
• Grosset C, Chen CY, Xu N et al. A mechanism for translationally coupled mRNA turnover: interaction between the poly(A) tail and a c-fos RNA coding determinant via a protein complex. Cell. 2000-09-29 [PMID: 11051545] (WB, Mouse)

Review for PABP Antibody (NB120-6125) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human, Oncorhynchus mykiss, Rainbow trout.

reviewed by: Verified Customer WB Human, Oncorhynchus mykiss, Rainbow trout 05/02/2017

Summary

• Application: Western Blot
• Sample Tested: HFF-1 cells total lysate,RTG-2 cells total lysate
• Species: Human, Oncorhynchus mykiss, Rainbow trout
• Lot: A-1

Bioinformatics

• Gene Symbol: PABPC1
• Entrez:
  • 26986(Human)
• Uniprot:
  • P11940(Human)