Human BTNL8 Antibody

Catalog #: MAB113641 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB113641-100
MAB113641-SP

Detection of Human BTNL8 by Western Blot.

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Human BTNL8 Antibody Summary

Species Reactivity

Human

Specificity

Detects human BTNL8 in direct ELISA

Source

Monoclonal Mouse IgG_2B Clone # 1062028

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Synthetic peptide Fc-cleaved human BTNL8

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

HEK293T human embryonic kidney cells transfected with human BTNL8.

Immunohistochemistry

5-25 µg/mL

Immersion fixed paraffin-embedded sections of Human Colon

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human BTNL8 by Western Blot. Western blot shows lysates of HEK293T human embryonic kidney cells transfected with human BTNL8. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human BTNL8 Monoclonal Antibody (Catalog # MAB113641) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for BTNL8 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Immunohistochemistry

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Detection of BTNL8 in Human Colon. BTNL8 was detected in immersion fixed paraffin-embedded sections of Human Colon using Mouse Anti-Human BTNL8 Monoclonal Antibody (Catalog # MAB113641) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane in lymphocytes in lamina propria. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: BTNL8

Butyrophilin-like 8 (BTNL8) is a member of the BTN/MOG Ig-superfamily and functions as a negative regulator of immune cell activation (1). Human BTNL8 is a 500 amino acid (aa) type I transmembrane glycoprotein that contains a signal peptide followed by an extracellular domain (ECD), a transmembrane region and a short cytoplasmic domain (2). The ECD of human BTNL8 shares 88% sequence identity with the ECD of mouse BTNL8. BTNL8 has two alternatively spliced forms: B7-like and BTN-like. Both isoforms of BTNL8 are expressed in a range of human tissues (3). The complete immunological function of BTNL molecules is only beginning to emerge. BTNL8 has been shown to be important in initiation of primary immune responses, suggesting a role in priming of naïve T lymphocytes (3). Down-regulation of BNTL8 mRNA levels has been associated with ulcerative colitis and colon cancer (4). BTNL8 are expressed in colon, lung, testis and neutrophils, and its expression is significantly decreased in ulcerative colitis, colonic tumors as compared to unaffected tissue (4). Soluble BTNL8-Fc fusion protein binds to resting, but not activated T cells. In vitro, BTNL8 co-stimulates T cell proliferation and cytokine production. In vivo injections of BTNL8-Fc significantly increases production of Ag‑specific IgG during the primary but not the secondary immune response (3).

References