Anti-MICA Polyclonal Antibody is a Rabbit antibody targeting MICA. Anti-MICA Polyclonal Antibody can be used in FCM,IF,IHC-Fr,IHC-P,WB.

IgG

Human,Mouse

1. Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-MICA/MHC I a Polyclonal Antibody, Unconjugated (TMAB-01136) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: mouse intestine tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-MICA/MHC I a Polyclonal Antibody, Unconjugated (TMAB-01136) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Tissue/cell: mouse lung tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-MICA/MHC I a Polyclonal Antibody, Unconjugated (TMAB-01136) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
4. Blank control (blue line): A431 (blue).
Primary Antibody (green line): Rabbit Anti-MICA antibody (TMAB-01136)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1 μg/test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
5. Sample:
Lane 1: Mouse Large intestine tissue lysates
Lane 2: Mouse Lung tissue lysates
Lane 3: Mouse Kidney tissue lysates
Lane 4: Mouse Liver tissue lysates
Primary: Anti-MICA (TMAB-01136) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 43 kDa
Observed band size: 60 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human MICA

Human

MICA

MHC class I chain-related protein A

Class I,NK Cells

Seems to have no role in antigen presentation. Acts as a stress-induced self-antigen that is recognized by gamma delta T-cells. Ligand for the KLRK1/NKG2D receptor. Binding to KLRK1 leads to cell lysis.