Recombinant Full Length Human KRT19 Protein, His-tagged

• Cat.No.: KRT19-7239H
• Product Overview: Recombinant Full Length Human KRT19 Protein with His tag was expressed in E. coli.

Specification

• Species: Human
• Source: E.coli
• Tag: His
• Protein Length: 1-400 aa
• Description: The protein encoded by this gene is a member of the keratin family. The keratins are intermediate filament proteins responsible for the structural integrity of epithelial cells and are subdivided into cytokeratins and hair keratins. The type I cytokeratins consist of acidic proteins which are arranged in pairs of heterotypic keratin chains. Unlike its related family members, this smallest known acidic cytokeratin is not paired with a basic cytokeratin in epithelial cells. It is specifically expressed in the periderm, the transiently superficial layer that envelopes the developing epidermis. The type I cytokeratins are clustered in a region of chromosome 17q12-q21.
• Tag: His
• Molecular Mass: 47 kDa
• AA Sequence: MGSSHHHHHHSSGLVPRGSHMGSMTSYSYRQSSATSSFGGLGGGSVRFGPGVAFRAPSIHGGSGGRGVSVSSARFVSSSSSGAYGGGYGGVLTASDGLLAGNEKLTMQNLNDRLASYLDKVRALEAANGELEVKIRDWYQKQGPGPSRDYSHYYTTIQDLRDKILGATIENSRIVLQIDNARLAADDFRTKFETEQALRMSVEADINGLRRVLDELTLARTDLEMQIEGLKEELAYLKKNHEEEISTLRGQVGGQVSVEVDSAPGTDLAKILSDMRSQYEVMAEQNRKDAEAWFTSRTEELNREVAGHTEQLQMSRSEVTDLRRTLQGLEIELQSQLSMKAALEDTLAETEARFGAQLAHIQALISGIEAQLGDVRADSERQNQEYQRLMDIKSRLEQEIATYRSLLEGQEDHYNNLSASKVL
• Purity: >90% by SDS-PAGE
• Storage: Store it under sterile conditions at -20 to -80 °C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Concentration: 1mg/mL by BCA
• Storage Buffer: Sterile PBS, pH7.4
• Publications:
  Pancreatic cancer cells assemble a CXCL12-keratin 19 coating to resist immunotherapy (2020)
  Carcinomas assemble a filamentous CXCL12–keratin-19 coating that suppresses T cell–mediated immune attack (2022)

Gene Information

• Gene Name: KRT19 keratin 19 [ Homo sapiens (human) ]
• Official Symbol: KRT19
• Synonyms: KRT19; keratin 19; keratin, type I cytoskeletal 19; 40 kDa keratin intermediate filament; CK19; cytokeratin 19; K1CS; K19; keratin; type I cytoskeletal 19; type I; 40 kd; MGC15366; CK-19; keratin-19; cytokeratin-19; keratin, type I, 40-kd; 40-kDa keratin intermediate filament
• Gene ID: 3880
• mRNA Refseq: NM_002276
• Protein Refseq: NP_002267
• MIM: 148020
• UniProt ID: P08727

Citation

Pancreatic cancer cells assemble a CXCL12-keratin 19 coating to resist immunotherapy

Journal: bioRxiv    Data: 2020/9/4

Authors: Wang Zhikai, Yan Ran, Fearon Douglas T.

Article Snippet:PrePrint: The buffer of commercially available recombinant human KRT19 protein (Creative BioMart, KRT19-7239H) was changed to protein binding buffer (25 mM Tris, pH 7.5, 100 mM NaCl, 1% Triton X-100), using Zeba spin desalting columns (Thermo, 89882).. For protein binding, biotinylated recombinant human CXCL12 (Chemotactics, B-CXCL12) or CXCL8 (Chemotactics, B-CXCL8) was incubated with KRT19 at 4°C for 2 hours.For protein binding, biotinylated recombinant human CXCL12 (Chemotactics, B-CXCL12) or CXCL8 (Chemotactics, B-CXCL8) was incubated with KRT19 at 4°C for 2 hours.

( A ) Sections of freshly resected human PDA and CRC were stained with fluorochrome-conjugated antibodies to KRT19 and CXCL12. Scale bar, 100 μm. ( B ) SDS-eluates of samples of the resected PDA and CRC were 5 subjected to SDS-PAGE and immunoblotting with antibodies to CXCL12 and KRT19. ( C ) The proteins that were immunoprecipitated by control IgG or anti-CXCL12 IgG from the pooled SDS-eluates of the four tumors were subjected to SDS-PAGE and immunoblotting with anti-CXCL12 and anti-KRT19 antibodies. ( D ) Streptavidin-beads bearing CXCL12-biotin were incubated with increasing concentrations of KRT19. Bound KRT19 was detected by SDS-PAGE 10 and immunoblotting with anti-KRT19 antibody. The apparent affinity with which CXCL12 and KRT19 interacted was calculated. ( E ) Anti-KRT19 antibody/protein G beads were incubated at 4°C with combinations of KRT19, CXCL12 and TGM2, in the presence or absence of Ca 2+ . The bound proteins were detected by SDS-PAGE and immunoblotting with the indicated antibodies. ( F ) Streptavidin beads bearing preformed complexes of biotinylated CXCL12 and KRT19 were 15 incubated at 20°C for 15 min with TGM2 in the presence or absence of the TGM2 inhibitor, ERW1041E. The proteins were eluted from the beads and detected by SDS-PAGE and immunoblotting with antibodies to CXCL12 and KRT19.

( A - B ) Sections of FFPE human carcinomas and melanoma (A) and frozen human carcinomas (B) were stained with fluorescent antibodies to KRT19 and CXCL12. Scale bars, 100 μm. ( C ) The NP-40 eluates of the four human tumors were subjected to SDS-PAGE and immunoblotting with anti-CXCL12 or anti-KRT19 antibodies. The SDS eluate of patient 2 was applied as a positive control.

( A ) A frozen section of mouse autochthonous KPC tumor was stained with fluorescent antibodies to KRT19 and CXCL12. Scale bar, 100 μm. ( B ) Sequential detergent eluates of a mouse subcutaneous PDA were subjected to SDS-PAGE and immunoblotting with anti-KRT19 or anti-CXCL12 antibody. ( C ) The proteins that were immunoprecipitated by control IgG or anti-CXCL12 antibody from the SDS-eluate of a mouse subcutaneous PDA were subjected to SDS-PAGE and immunoblotting with anti-CXCL12 or anti-KRT19 antibodies. ( D ) The mRNAs of Cxcl12, Pdgfra (indicating CAFs) and Krt19 (PDA cells) were detected by RNA fluorescent in situ hybridization (FISH) in sections of a mouse subcutaneous PDA. The pie chart shows relative proportions of the cells that are Pdgfra + , or Cxcl12 + or double positive. Scale bar, 50 μm. ( E ) 30 μm-thick sections of a mouse subcutaneous PDA were permeabilized with Triton X-100, or not, were stained with fluorescent antibodies to KRT19, CXCL12, and tubulin. Scale bars, 10 μm. ( F ) Intact, DAPI-excluding dissociated cells from a mouse subcutaneous PDA tumor that had been stained with fluorescent antibodies to CXCL12, KRT19 and CD45, were assessed by flow cytometry.