Recombinant Human OLFM4 protein(Met1-Gln510), His-tagged

• Cat.No.: OLFM4-3222H
• Product Overview: Recombinant Human OLFM4 (NP_006409.3) (Met 1-Gln 510) was expressed in HEK293, fused with a polyhistidine tag at the C-terminus.

Specification

• Species: Human
• Source: HEK293
• Tag: His
• Protein Length: 1-510 a.a.
• Form: Lyophilized from sterile PBS, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
• Molecular Mass: The recombinant human OLFM4 consists of 501 amino acids and predictes a molecular mass of 56.6 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh OLFM4 is approximately 65-70 kDa due to different glycosylation.
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method
• Purity: > 92 % as determined by SDS-PAGE
• Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C. Store it under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Gene Information

• Gene Name: OLFM4 olfactomedin 4 [ Homo sapiens ]
• Official Symbol: OLFM4
• Synonyms: OLFM4; olfactomedin 4; olfactomedin-4; GC1; GW112; OlfD; antiapoptotic protein GW112; G-CSF-stimulated clone 1 protein; OLM4; hGC-1; hOLfD; UNQ362; bA209J19.1; KIAA4294
• Gene ID: 10562
• mRNA Refseq: NM_006418
• Protein Refseq: NP_006409
• MIM: 614061
• UniProt ID: Q6UX06

Citation

The olfactomedin-4 positive neutrophil has a role in murine intestinal ischemia/reperfusion injury

Journal: The FASEB Journal    PubMed ID: 31593636    Data: 2020/11/25

Authors: Nick C. Levinsky, Jaya Mallela, Matthew N. Alder

Article Snippet:To evaluate whether secreted OLFM4 may have an effect on surrounding inflammatory cells, murine RAW264 cells were grown in tissue culture with DMEM with 10% fetal bovine serum and plated in 6-well plates at 1 × 10 6 cells/well until reaching 50–60% confluency.To evaluate whether secreted OLFM4 may have an effect on surrounding inflammatory cells, murine RAW264 cells were grown in tissue culture with DMEM with 10% fetal bovine serum and plated in 6-well plates at 1 × 10 6 cells/well until reaching 50–60% confluency.. They were then stimulated overnight with 10–100 ng/ml of recombinant murine OLFM4 (Creative BioMart, Shirley, NY, USA) at 37°C.. Control treatments included medium alone or 1 μg/ml of LPS.Control treatments included medium alone or 1 μg/ml of LPS.

OLFM4 null mice are protected from intestinal IR injury. A) Kaplan-Meier survival curve demonstrating mortality at 7 d following IR injury. OLFM4 null mice have a significant survival benefit following intestinal IR injury compared with WT mice undergoing IR (33 vs. 68%, P = 0.028). B) Assay of intestinal permeability by plasma levels of FITC-dextran at 4 h following reperfusion. WT mice have significantly higher plasma FITC-dextran concentration at 4 h after reperfusion compared with OLFM4 null mice (414 ± 414 vs. 162 ng/ml ± 147, P = 0.031; Wilcoxon rank sum).

OLFM4 null mice have less histologic evidence of injury. A) Representative light microscopy images of mouse intestinal tissue stained with hematoxylin and eosin. Healthy control tissue for WT (top left) and OLFM4 null (bottom left) are demonstrated. Following IR injury, WT mice have severe injury up to complete loss of mucosal architecture (top right), whereas OLFM4 null mice have minimal visible evidence of IR injury (bottom right). B) Mean injury scores of WT and OLFM4 null mice after injury as graded with the Chiu score by 4 blinded observers. WT mice had significantly higher mean injury scores compared with OLFM4 null mice (3.1 ± 1.5 vs. 1.3 ± 1.1, P = 0.019; Student’s t test). Furthermore, OLFM4 null mice had similar mean injury scores as healthy controls.

Representative immunohistochemical sections stained for OLM4 in mouse intestine. A) In healthy control WT mice, OLFM4 is located in the bases of the crypts. B) Following IR injury in WT mice, OLFM4 expression appears to extend further up the crypts, and OLFM4 is found secreted into the lumen (double arrow). OLFM4-positive neutrophils are also noted infiltrating the base of the crypts after IR injury (single arrows). C) OLFM4 null mice after IR injury shown for comparison, where OLFM4 is not present. D) Western blotting of neutrophil cell pellets or culture medium following stimulation with 100 nM PMA. Unstimulated WT neutrophil pellets have high levels of OLFM4 compared with stimulated. Culture medium from stimulated WT neutrophils has OLFM4, whereas there is no OLFM4 in the medium from unstimulated neutrophils. There is no OLFM4 in the null pellets or medium.