RALY Monoclonal antibody

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  WB analysis of LNCaP using 68011-1-Ig

  LNCaP cells were subjected to SDS PAGE followed by western blot with 68011-1-Ig (RALY antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with Lamin B1 Monoclonal antibody (66095-1-Ig) as loading control.

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  IHC staining of human stomach cancer using 68011-1-Ig

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 68011-1-Ig (RALY antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human stomach cancer using 68011-1-Ig

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 68011-1-Ig (RALY antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of A431 using 68011-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed A431 cells using RALY antibody (68011-1-Ig, Clone: 2B10C3 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  FC experiment of A431 using 68011-1-Ig

  1X10^6 A431 cells were intracellularly stained with 0.4 ug Anti-Human RALY (68011-1-Ig, Clone:2B10C3) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

RALY Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Cat No. 68011-1-Ig

Clone No.2B10C3

Publications(1)

Host / Isotype

Mouse / IgG1

Reactivity

human

Applications

WB, IHC, IF/ICC, FC (Intra), CoIP, ELISA

Synonyms

2B10C3, Autoantigen p542, Heterogeneous nuclear ribonucleoprotein C-like 2, hnRNP associated with lethal yellow protein homolog, hnRNP core protein C-like 2

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Tested Applications

• Positive WB detected in: LNCaP cells, HeLa cells, HepG2 cells, Jurkat cells, K-562 cells
• Positive IHC detected in: human stomach cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: A431 cells
• Positive FC (Intra) detected in: A431 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 1 publications below
• CoIP: See 1 publications below

Product Information

68011-1-Ig targets RALY in WB, IHC, IF/ICC, FC (Intra), CoIP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag12250

  Product name: Recombinant human RALY protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 1-307 aa of BC103753

  Sequence: MSLKLQASNVTNKNDPKSINSRVFIGNLNTALVKKSDVETIFSKYGRVAGCSVHKGYAFVQYSNERHARAAVLGENGRVLAGQTLDINMAGEPKPDRPKGLKRAASAIYSGYIFDYDYYRDDFYDRLFDYRGRLSPVPVPRAVPVKRPRVTVPLVRRVKTNVPVKLFARSTAVTTSSAKIKLKSSELQAIKTELTQIKSNIDALLSRLEQIAAEQKANPDGKKKGDGGGASGGGGGGGGSGGGGSGGGGGGGSSRPPAPQENTTSEAGLPQGEARTRDDGDEEGLLTHSEEELEHSQDTDADDGALQ

• Full Name: RNA binding protein, autoantigenic (hnRNP-associated with lethal yellow homolog (mouse))
• Calculated Molecular Weight: 307 aa, 33 kDa
• Observed Molecular Weight: 30-37 kDa
• GenBank Accession Number: BC103753
• Gene Symbol: RALY
• Gene ID (NCBI): 22913
• RRID: AB_2918758
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: Q9UKM9
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

RNA-binding protein RALY is a protein that in humans is encoded by the RALY gene. In infectious mononucleosis, anti-EBNA-1 antibodies are produced which cross-react with multiple normal human proteins. The cross-reactivity is due to anti-gly/ala antibodies that cross-react with host proteins containing configurations like those in the EBNA-1 repeat. One such antigen is RALY which is a member of the heterogeneous nuclear ribonucleoprotein gene family. RALYhas two isoforms with the molecular weight of 30 kDa and 32 kDa, and it has many modification sites, which made it shows at 30-37 kDa by Western Blot.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	WB,CoIP	Int J Biol Sci; O-GlcNAcylated RALY Contributes to Hepatocellular Carcinoma Cells Proliferation by Regulating USP22 mRNA Nuclear Export; Authors - Shiwei Liu ; KD Validated