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MIF Recombinant Rabbit Monoclonal Antibody [PSH06-49] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751079

MIF Recombinant Rabbit Monoclonal Antibody [PSH06-49] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

MIF Recombinant Rabbit Monoclonal Antibody [PSH06-49] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human MIF aa 1-115.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 12 kDa

Positive Control

Jurkat cell lysate, THP-1 cell lysate, HL-60 cell lysate, U-87 MG cell lysate, HEK-293 cell lysate, A549 cell lysate, HepG2 cell lysate, HUVEC cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Jurkat.

Conjugation

unconjugated

Clone Number

PSH06-49

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:10,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

Macrophage migration inhibitory factor (MIF), also known as glycosylation-inhibiting factor (GIF), L-dopachrome isomerase, or phenylpyruvate tautomerase is a protein that in humans is encoded by the MIF gene. MIF is an important regulator of innate immunity. The MIF protein superfamily also includes a second member with functionally related properties, the D-dopachrome tautomerase (D-DT). CD74 is a surface receptor for MIF. Bacterial antigens stimulate white blood cells to release MIF into the blood stream. The circulating MIF binds to CD74 on other immune cells to trigger an acute immune response. Hence, MIF is classified as an inflammatory cytokine. Furthermore, glucocorticoids also stimulate white blood cells to release MIF and hence MIF partially counteracts the inhibitory effects that glucocorticoids have on the immune system. Finally trauma activates the anterior pituitary gland to release MIF.

Background References

1. Sumaiya K et al. Macrophage migration inhibitory factor (MIF): A multifaceted cytokine regulated by genetic and physiological strategies. Pharmacol Ther. 2022 May

2. Matejuk A et al. MIF contribution to progressive brain diseases. J Neuroinflammation. 2024 Jan

Subcellular Location

Secreted, Cytoplasm.

UNIPROT

SwissProt: P14174 Human

SwissProt: P34884 Mouse

SwissProt: P30904 Rat

Synonyms

GIF antibody

GLIF antibody

Glycosylation inhibiting factor antibody

Glycosylation-inhibiting factor antibody

L-dopachrome isomerase antibody

L-dopachrome tautomerase antibody

Macrophage migration inhibitory factor (glycosylation-inhibiting factor) antibody

Macrophage migration inhibitory factor antibody

MIF antibody

MIF protein antibody

Expand

Images

  • Western blot analysis of MIF on different lysates with Rabbit anti-MIF antibody (<a href="/products/HA751079" style="font-weight: bold;text-decoration: underline;">HA751079</a>) at 1/2,000 dilution.<br /><br />Lane 1: Jurkat cell lysate (20 µg/Lane)<br />Lane 2: THP-1 cell lysate (20 µg/Lane)<br />Lane 3: HL-60 cell lysate (20 µg/Lane)<br />Lane 4: U-87 MG cell lysate (20 µg/Lane)<br />Lane 5: HEK-293 cell lysate (20 µg/Lane)<br />Lane 6: A549 cell lysate (20 µg/Lane)<br />Lane 7: HepG2 cell lysate (20 µg/Lane)<br />Lane 8: HUVEC cell lysate (20 µg/Lane)<br />Lane 9: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 10: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 11: C6 cell lysate (20 µg/Lane)<br />Lane 12: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 13: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 12 kDa<br />Observed band size: 12 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751079" style="font-weight: bold;text-decoration: underline;">HA751079</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of MIF on different lysates with Rabbit anti-MIF antibody (HA751079) at 1/2,000 dilution.

    Lane 1: Jurkat cell lysate (20 µg/Lane)
    Lane 2: THP-1 cell lysate (20 µg/Lane)
    Lane 3: HL-60 cell lysate (20 µg/Lane)
    Lane 4: U-87 MG cell lysate (20 µg/Lane)
    Lane 5: HEK-293 cell lysate (20 µg/Lane)
    Lane 6: A549 cell lysate (20 µg/Lane)
    Lane 7: HepG2 cell lysate (20 µg/Lane)
    Lane 8: HUVEC cell lysate (20 µg/Lane)
    Lane 9: Neuro-2a cell lysate (20 µg/Lane)
    Lane 10: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 11: C6 cell lysate (20 µg/Lane)
    Lane 12: Mouse brain tissue lysate (40 µg/Lane)
    Lane 13: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 12 kDa
    Observed band size: 12 kDa

    Exposure time: 6 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751079) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of Jurkat cells labeling MIF with Rabbit anti-MIF antibody (<a href="/products/HA751079" style="font-weight: bold;text-decoration: underline;">HA751079</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MIF antibody (<a href="/products/HA751079" style="font-weight: bold;text-decoration: underline;">HA751079</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Jurkat cells labeling MIF with Rabbit anti-MIF antibody (HA751079) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MIF antibody (HA751079) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of MIF on different lysates with Rabbit anti-MIF antibody (<a href="/products/HA751079" style="font-weight: bold;text-decoration: underline;">HA751079</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-MIF KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 12 kDa<br />Observed band size: 12 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751079" style="font-weight: bold;text-decoration: underline;">HA751079</a>) at 1/2,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of MIF on different lysates with Rabbit anti-MIF antibody (HA751079) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-MIF KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 12 kDa
    Observed band size: 12 kDa

    Exposure time: 6 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751079) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Flow cytometric analysis of Jurkat cells labeling MIF.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751079" style="font-weight: bold;text-decoration: underline;">HA751079</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Jurkat cells labeling MIF.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751079, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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