Human/Mouse/Rat MARCH8 Antibody

Name: Human/Mouse/Rat MARCH8 Antibody
Brand: R&D Systems
SKU: MAB6969

Catalog #: MAB6969 Datasheet / COA / SDS

Discontinued Product

MAB6969 has been discontinued.

View all MARCH8 products.

Detection of Human, Mouse, and Rat MARCH8 by Western Blot.

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All MARCH8 Antibodies

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat MARCH8 Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human MARCH8 in direct ELISAs. In Western blots, no cross-reactivity with recombinant human (rh) MARCH-1 or rhGRAIL is observed.

Source

Monoclonal Mouse IgG_2B Clone # 710424

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human MARCH8
Gln218-Val291
Accession # Q5T0T0

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.

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

See below

Immunocytochemistry

8-25 µg/mL

See below

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, Mouse, and Rat MARCH8 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, K562 human chronic myelogenous leukemia cell line, L1.2 mouse pro-B cell line, and Y3-Ag rat myeloid cell line. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human/Mouse/Rat MARCH8 Monoclonal Antibody (Catalog # MAB6969) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for MARCH8 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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MARCH8 in Human Dendritic Cells. MARCH8 was detected in immersion fixed immature human dendritic cells using Mouse Anti-Human/Mouse/Rat MARCH8 Monoclonal Antibody (Catalog # MAB6969) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

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: MARCH8

MARCH8 (membrane-associated RING-CH VIII), also called MIR (modulator of immune recognition), or RNF178 (ring finger 178), is a 33 kDa two-pass transmembrane protein of lysosomal and early endosomal membranes in the MARCH family. MARCH proteins are E3 ubiquitin ligases that promote endocytosis and degradation of their target proteins. MARCH8 ubiquitinates B7-2/CD86 and probably Fas, Tfr and MHC class II proteins such as HLA-A2.1. Within the cytoplasmic C-terminal region used as an immunogen, human MARCH8 shares 87% amino acid identity with mouse and rat MARCH8.

Long Name

Membrane Associated Ring Finger (C3HC4) 8

Entrez Gene IDs

220972 (Human); 71779 (Mouse); 312656 (Rat)

Alternate Names

Cellular modulator of immune recognition; c-MIR; EC 6.3.2; EC 6.3.2.-; MARCH8; MARCH-VIII; MARCH-VIIIcellular modulator of immune recognition; membrane-associated ring finger (C3HC4) 8; Membrane-associated RING finger protein 8; Membrane-associated RING-CH protein VIII; MIR; RING finger protein 178; RNF178