Antibody data
- Antibody Data
- Antigen structure
- References [4]
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- Validations
- Western blot [5]
- ELISA [1]
- Immunocytochemistry [2]
- Immunohistochemistry [2]
- Other assay [6]
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- Product number
- PA5-27343 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MIF Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: Molt-4, mouse brain.
- Concentration
- 1 mg/mL
Submitted references Neutrophils delay repair process in Wallerian degeneration by releasing NETs outside the parenchyma.
Macrophage Migration Inhibitory Factor (MIF) and Its Homologue d-Dopachrome Tautomerase (DDT) Inversely Correlate with Inflammation in Discoid Lupus Erythematosus.
CD74 Signaling Links Inflammation to Intestinal Epithelial Cell Regeneration and Promotes Mucosal Healing.
Defense and protection mechanisms in lung exposed to asbestiform fiber: the role of macrophage migration inhibitory factor and heme oxygenase-1.
Yamamoto Y, Kadoya K, Terkawi MA, Endo T, Konno K, Watanabe M, Ichihara S, Hara A, Kaneko K, Iwasaki N, Ishijima M
Life science alliance 2022 Oct;5(10)
Life science alliance 2022 Oct;5(10)
Macrophage Migration Inhibitory Factor (MIF) and Its Homologue d-Dopachrome Tautomerase (DDT) Inversely Correlate with Inflammation in Discoid Lupus Erythematosus.
Caltabiano R, De Pasquale R, Piombino E, Campo G, Nicoletti F, Cavalli E, Mangano K, Fagone P
Molecules (Basel, Switzerland) 2021 Jan 1;26(1)
Molecules (Basel, Switzerland) 2021 Jan 1;26(1)
CD74 Signaling Links Inflammation to Intestinal Epithelial Cell Regeneration and Promotes Mucosal Healing.
Farr L, Ghosh S, Jiang N, Watanabe K, Parlak M, Bucala R, Moonah S
Cellular and molecular gastroenterology and hepatology 2020;10(1):101-112
Cellular and molecular gastroenterology and hepatology 2020;10(1):101-112
Defense and protection mechanisms in lung exposed to asbestiform fiber: the role of macrophage migration inhibitory factor and heme oxygenase-1.
Loreto C, Caltabiano R, Graziano ACE, Castorina S, Lombardo C, Filetti V, Vitale E, Rapisarda G, Cardile V, Ledda C, Rapisarda V
European journal of histochemistry : EJH 2020 Apr 16;64(2)
European journal of histochemistry : EJH 2020 Apr 16;64(2)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MIF using 30 µg of NIH-3T3 lysate. Samples were loaded onto a 15% SDS-PAGE gel and probed with a MIF polyclonal antibody (Product # PA5-27343) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis MIF was performed by loading 30 µg of HEK293T whole cell lysate per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat dry milk in TBST for 1 hour at room temperature. The membrane was probed with a MIF polyclonal antibody (Product # PA5-27343) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MIF was performed by separating 50 µg of various tissue extracts by 15% SDS-PAGE. Proteins were transferred to a membrane and probed with a MIF Polyclonal Antibody (Product # PA5-27343) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using MIF Polyclonal Antibody (Product # PA5-27343). Sample (30 µg of whole cell lysate). A: Molt-4.15% SDS PAGE. MIF Polyclonal Antibody (Product # PA5-27343) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using Anti-MIF Polyclonal Antibody (Product # PA5-27343) and a 15 kDa band corresponding to Macrophage migration inhibitory factor was observed across cell lines tested and was upregulated in U-87 MG upon VEGF treatment. Whole cell extracts (30 µg lysate) of THP-1 (Lane 1), U-937 (Lane 2), U-87 MG (Lane 3), U-87 MG treated with VEGF (100 ng/mL for 48hrs) (Lane 4), Jurkat (Lane 5), RAW 264.7 (Lane 6), HEK-293 (Lane 7) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Uncharacterized bands were observed at 30 and 60 kDa.
Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ELISA detection of MIF using MIF Polyclonal Antibody (Product # PA5-27343) for capture at a concentration of 5 µg/mL and a MIF Polyclonal Antibody for detection at a concentration of 1.5 µg/mL.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of MIF in methanol-fixed HeLa cells using a MIF polyclonal antibody (Product # PA5-27343) at a 1:200 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of 14-3-3 zeta (green) HEK293T cells. Cells fixed with 4% formaldehyde fixed were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with a 14-3-3 zeta ppolyclonal antibody (Product # PA5-27317) at a dilution of 1:100 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with fluorophore-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification. Data courtesy of the Innovators Program.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry (Paraffin) analysis of MIF was performed in paraffin-embedded mouse brain tissue using MIF Polyclonal Antibody (Product # PA5-27343) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry (Paraffin) analysis of MIF was performed in paraffin-embedded rat brain tissue using MIF Polyclonal Antibody (Product # PA5-27343) at a dilution of 1:500.
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Figure 1 Immunohistochemistry analysis of MIF expression in normal skin biopsies. High levels of MIF can be detected at the basal layer of the epidermis (red arrow) ( A ) and in the cutaneous appendages (eccrine glands and sebocytes) (black arrow) ( B ). Immunohistochemistry-positive staining was defined as the presence of brown chromogen detection within the cytoplasm. Representative microphotographs are shown.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Figure 2 Immunohistochemistry analysis of MIF expression in DLE skin biopsies. Thirty-seven patients with DLE were recruited for this study. Biopsies were performed after a six-month wash-out period from topical treatments. Hematoxylin and eosin staining was used for histopathological evaluation. Representative microphotographs are shown. ( A - D ) In the presence of a high inflammatory score (2+; 3+), MIF low expression (0; 1+) was observed in the epidermis (one arrow), appendages (two arrows), and inflammatory infiltrate (three arrows). (( A , B ): patient no. 1; ( C , D ): patient no. 2). ( E - H ) In the presence of a low inflammatory score (0; 1+), MIF high expression (2+; 3+) was observed in the epidermis (one arrow), appendages (two arrows), and inflammatory infiltrate (three arrows). Immunohistochemistry positive staining was defined as the presence of brown chromogen detection within the cytoplasm. ( E , F ): patient no. 3; ( G , H ): patient no. 4).
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- Invitrogen Antibodies (provider)
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- Figure 1. A1) MIF IHC section of non-exposed sheep (magnification 200x). A2) MIF IHC section of exposed sheep; the fibrotic interstitium showed a strong scattered immunoreaction; green arrow indicates the intra-parenchymal stroma around a bronchiolar structure, yellow arrow shows MIF macrophages immunodetection while red arrow indicates FE deposit fibers (magnification 200x). A3) MIF immunostaining software image analysis of panel A2, in which mainly a high immunostained area (red color) was detected (magnification 200x). B1) HO-1 IHC section of non-exposed sheep (magnification 200x). B2) HO-1 IHC section of exposed sheep; in the lung fibrotic tissue, a strong and widespread immunostaining was demonstrated throughout the interstitium (black arrow) and bronchiolar structures (red arrow) (magnification 200x). B3) HO-1 immunostaining software image analysis of panel B2, in which mainly a high immunostained area (red color) was detected (magnification 200x).
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- Figure 2. A) Higher magnification of MIF immunohistochemical expression in exposed sheep section (magnification 400x). B) Higher magnification of HO-1 immunohistochemical expression in exposed sheep section (magnification 400x).
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- Experimental details
- Figure 4. HO-1 and MIF protein levels in human primary lung fibroblasts unexposed or FE-exposed for 72 h. Representative immunoblotting of HO-1 and MIF expressions (A). Results of three independent immunoblots are represented as percentage of HO-1 (B) and MIF (C) proteins with respect to untreated cells (*P
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- Figure 8. Migration inhibitory factor (MIF) secreted from neutrophils promotes NET formation in rats. (A) Representative images of triple immunolabeling of longitudinal sections with MIF, Ly6G, and DAPI. Left is intact nerve, and other images were acquired at 20 mm distal to the injury site 12 h after injury. Dashed lines indicate the border between the epineurium and the parenchyma. The images marked with * is a high-magnification image of the boxed area. Arrows indicate neutrophils expressing MIF. Neutrophils at the epineurium expressed MIF. Scale bars: 10 mum in * images and 20 mum in all other images. (B) Schematic illustration of an experimental method to inhibit MIF at the epineurium in Wallerian degeneration. The nerve of the region of Wallerian degeneration was wrapped with collagen sheets containing iso-1 or control. (C) Representative images of IgG immunolabeling at 20 mm distal to the injury site. No IgG immunoreactivity was detected at the parenchyma in either group. Scale bar: 500 mum. (D) Representative images of immunolabeling with CitH3, MPO, and DAPI at 20 mm distal to the injury site 12 h after injury. Dashed lines indicate the border of the epineurium and the parenchyma. Images marked with * are high-magnification images of boxed areas in the epineurium. Arrows indicate triple immunolabeled NETs. Treatment with iso-1 dramatically inhibited NET formation. Scale bar: 10 mum. (E) Quantification of the NET formation detected by triple labeling with the CitH3, M