Palladin Antibody (1E6)

Knockdown Validated: Palladin Antibody (1E6) [NBP1-25959] - C2C12 cells were transfected with shRNA targeting palladin or nontargeting shRNA as a control. Stable transfectants were submitted to a differentiation medium and followed for 5 days in culture (D0 to D5). Myogenic differentiation was assessed at the indicated time points using RNA/protein analysis and immunofluorescence staining. Immunofluorescence images of stable transfectants labeled with palladin antibody (green) and F-actin (red). Blue is DAPI-stained nuclei. Scale bar is 10 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0124762) licensed under a CC-BY license.

Western Blot: Palladin Antibody (1E6) [NBP1-25959] - MDA-MB-231 lysate.

Knockdown Validated: Palladin Antibody (1E6) [NBP1-25959] - Western blots analysis of C2C12 cells labeled with a monoclonal antibody against 90-kDa palladin or a polyclonal antibody against 140- and 200-kDa palladin, MyHC, myogenin, and MLC. The blots clearly show the decrease in palladin expression concomitant with the elevation of differentiation markers. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0124762) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Palladin Antibody (1E6) [NBP1-25959] - Immunofluorescence on a7r5 cells.

Western Blot: Palladin Antibody (1E6) [NBP1-25959] - Western blot analysis of HeLa (A), A431 (B), NIH3T3 (C), and PC-12 (D) cell line extracts using Palladin antibody at 1ug/ml.

Immunohistochemistry-Paraffin: Palladin Antibody (1E6) [NBP1-25959] - Palladin was detected in immersion fixed paraffin-embedded sections of human heart using Mouse Anti-Human Palladin (1E6) Monoclonal Antibody (Catalog # NBP1-25959) at 1:100 for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm in cardiomyocytes.

Flow Cytometry: Palladin Antibody (1E6) [NBP1-25959] - An intracellular stain was performed on HeLa cells with Palladin Antibody [1E6] NBP1-25959AF488 (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 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

• Reactivity: Hu, Mu, Rt, Ca, Ch
• Applications: WB, Flow, ICC/IF, IHC, IP, KD
• Clone: 1E6
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Palladin Antibody (1E6) Summary

• Immunogen: Endogenous Palladin immunoprecipitated from chicken embryo fibroblasts.
• Localization: Cytoplasm, cytoskeleton. Note: Localizes to stress fibers and Z-disks. Cell junction, focal adhesion. Cell projection, ruffle. Cell projection, lamellipodium.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: PALLD
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 5 ug/ml
  • Immunocytochemistry/Immunofluorescence 1:20-1:100
  • Immunohistochemistry 1:50-1:200
  • Immunohistochemistry-Paraffin 1:50-1:100
  • Immunoprecipitation 0.05 ml / 2 ml lysate
  • Knockdown Validated
  • Western Blot 1 ug/ml
• Application Notes: In WB a band is seen at ~85 kDa. 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.
• Theoretical MW: 85 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.

Reactivity Notes

Canine reactivity reported in scientific literature (PMID: 23451256).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine and 0.15M NaCl
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for Palladin Antibody (1E6)

• CGI151
• CGI-151
• FLJ22190
• FLJ38193
• KIAA0992FLJ39139
• MYN
• myoneurin
• Palladin
• palladin, cytoskeletal associated protein
• PALLD
• PNCA1
• Sarcoma antigen NY-SAR-77
• SIH002
• SIH002FLJ61376

Background

Palladin (PALLD) is an actin associated cytoskeletal protein which is widely expressed and found in stress fibers, focal adhesions, growth cones, Z-discs, and other actin-based subcellular structures. It belongs to a small gene family that includes the Z-disc proteins myopalladin and myotilin, all of which share similar Ig-like domains and interact with specific molecular partners: all three bind to alpha-actinin; in addition, palladin also binds to VASP and ezrin, myotilin binds to filamin and actin, and myopalladin also binds to nebulin and CARP. While myotilin and myopalladin are most abundant in skeletal and cardiac muscle, palladin is ubiquitously expressed in the organs of developing vertebrates. Palladin functions as a molecular scaffold as well as an actin cross linker, and its nuclear presence and ability to translocate to the nucleus upon stimulation reflects that palladin has signaling functions in excitation-transcription. It interacts with EPS8, LASP1, VASP, ACTN, ARGBP2, LPP, PFN1, SPIN90, SRC and EZR, and plays key role in smooth muscle differentiation, migration, contraction, embryonic development and in some pathological conditions including cardiovascular diseases as well as cancers. Palladin levels are typically upregulated actively migrating cells e.g. developing vertebrate embryos, in cells along a wound edge, and in metastatic cancer cells; and genetic variations in PALLD gene have been linked with susceptibility to pancreatic cancer type 1 (PNCA1) and myocardial infarction.

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 Palladin Antibody (NBP1-25959)(19)

Publications using NBP1-25959

• Mayer O, Bugis J, Kozlova D et al. Cytoskeletal Protein Palladin in Adult Gliomas Predicts Disease Incidence, Progression, and Prognosis Cancers 2022-10-19 [PMID: 36291914] (IF/IHC, Mouse)
• Davidson B, Bock Aj, Holth A, Nymoen Da Expression of palladin is associated with disease progression in metastatic high-grade serous carcinoma Cytopathology 2020-08-02 [PMID: 32741023] (IF/IHC, Human)
• Bhome R, Goh RW, Bullock MD et al. Exosomal microRNAs derived from colorectal cancer-associated fibroblasts: role in driving cancer progression Aging (Albany NY) 2017-12-28 [PMID: 29283887] (WB, Human)
• Li S, Liu C, Gu L et al. Autophagy protects cardiomyocytes from the myocardial ischaemia-reperfusion injury through the clearance of CLP36. Open Biol. 2016-08-01 [PMID: 27512143] (Mouse)
• Morishige N, Murata S, Nakamura Y et al. Coordinated Regulation of Palladin and alpha-Smooth Muscle Actin by Transforming Growth Factor-b in Human Corneal Fibroblasts. Invest. Ophthalmol. Vis. Sci. 1905-07-08 [PMID: 27367503] (ICC/IF, Human)
• Chang EH, Gasim AH, Kerber ML et al. Palladin is Upregulated in Kidney Disease and Contributes to Epithelial Cell Migration After Injury Sci Rep. 2015-01-09 [PMID: 25573828] (WB, Human, Mouse)
• van Steenbeek FG, Van den Bossche L, Grinwis GC et al. Aberrant Gene Expression in Dogs with Portosystemic Shunts. PLoS One 2013-01-01 [PMID: 23451256] (IHC-P, Canine)
• Dhanda A Morphological and functional characterization of host proteins during infections by actin-hijacking bacterial pathogens Thesis 2020-01-01 (ICC/IF, Human, Mammal)
  
  Details:
  Potorous tridactylus kidney
• Dhanda AS, Vogl AW, Albraiki SE et al. Palladin Compensates for the Arp2/3 Complex and Supports Actin Structures during Listeria Infections MBio 2018-04-10 [PMID: 29636431] (ICC/IF, WB, Human)
• Nguyen NU, Wang HV. Dual roles of palladin protein in in vitro myogenesis: inhibition of early induction but promotion of myotube maturation PLoS ONE 2015-04-16 [PMID: 25875253] (WB, Mouse)
• Stasiak M, Gawryś K, Popielarski M et al. Differential Quantitative Proteomics of Human Microvascular Endothelial Cells 1 by iTRAQ Reveals Palladin to be a New Biomarker During TGF-beta 1 Induced Endothelial Mesenchymal Transition Journal of Proteomics & Bioinformatics 2017-10-11 (WB, Human)
• Brentnall TA, Lai LA, Coleman J et al. Arousal of cancer-associated stroma: overexpression of palladin activates fibroblasts to promote tumor invasion. PLoS ONE 20127(1):e30219. [PMID: 22291919] (ICC/IF, IHC-P, Human)
• Goicoechea SM, Bednarski B, Stack C, Cowan DW, Volmar K, Thorne L, Cukierman E, Rustgi AK, Brentnall T, Hwang RF, McCulloch CA, Yeh JJ, Bentrem DJ, Hochwald SN, Hingorani SR, Kim HJ, Otey CA. Isoform-specific upregulation of palladin in human and murine pancreas tumors. PLoS One;5(4):e10347. 2010-04-26 [PMID: 20436683] (IHC-P, WB, Human)
• Goicoechea, S et al. Palladin contributes to invasive motility in human breast cancer cells. Oncogene;28(4):587-98. 2009-01-29 [PMID: 18978809] (WB, Human)
• Endlich N, Otey CA, Kriz W, Endlich K. Movement of stress fibers away from focal adhesions identifies focal adhesions as sites of stress fiber assembly in stationary cells. Cell Motil Cytoskeleton;64(12):966-76. 2007-12-01 [PMID: 17868136] (ICC/IF, Mouse)
• Wall ME, Rachlin A, Otey CA, Loboa EG. Human adipose-derived adult stem cells upregulate palladin during osteogenesis and in response to cyclic tensile strain. Am J Physiol Cell Physiol;293(5):C1532-8. 2007-11-01 [PMID: 17687002] (IHC-P, WB, Human)
• Goicoechea S, Arneman D, Disanza A, Garcia-Mata R, Scita G, Otey CA. Palladin binds to Eps8 and enhances the formation of dorsal ruffles and podosomes in vascular smooth muscle cells. J Cell Sci;119(Pt 16):3316-24. 2006-08-15 [PMID: 16868024] (IP, WB, Rat)
• Rachlin AS, Otey CA. Identification of palladin isoforms and characterization of an isoform-specific interaction between Lasp-1 and palladin. J Cell Sci;119(Pt 6):995-1004. 2006-03-15 [PMID: 16492705] (WB, Human, Mouse)
• Parast, M, Otey, A et al. Characterization of palladin, a novel protein localized to stress fibers and cell adhesions. J Cell Biol;150(3):643-56. 2000-08-07 [PMID: 10931874] (ICC/IF, IP, WB, Chicken, Mouse)

FAQs for Palladin Antibody (NBP1-25959). (Showing 1 - 1 of 1 FAQs).

1. What is the MW of the Palladin when run on a native gel?
   • The only QC experiments we have for this antibody in WB were run under reducing conditions, so we do not know the MW size when run on a native gel. I checked our WB publication list and did not see any evidence of the size of Palladin on a native gel. 

Bioinformatics

• Gene Symbol: PALLD
• Entrez:
  • 23022(Human)
• Uniprot:
  • Q8WX93()