MHC class I Recombinant Rabbit Monoclonal Antibody [JF10-38] - BSA and Azide free
Catalog# HA750347
MHC class I Recombinant Rabbit Monoclonal Antibody [JF10-38] - BSA and Azide free
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WB
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IF-Cell
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IHC-P
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FC
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IF-Tissue
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Human
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unconjugated
Overview
Product Name
MHC class I Recombinant Rabbit Monoclonal Antibody [JF10-38] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human HLA aa 263-299 / 365.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, FC, IF-Tissue
Target Molecular Weight
Predicted band size: 41 kDa
Positive Control
A431 cell lysate, HL-60 cell lysate, Jurkat cell lysate, Raji cell lysate, THP-1 cell lysate, HL-60, human colon cancer tissue, human liver cancer tissue, human tonsil tissue, human pancreas tissue.
Conjugation
unconjugated
Clone Number
JF10-38
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IF-Cell
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1:100
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IHC-P
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1:200-1:1:1,000
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FC
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1:1,000
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IF-Tissue
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1:200
Target
Function
Major histocompatibility complex (MHC) molecules, also designated human leukocyte antigen (HLA) molecules, are cell-surface receptors that bind foreign peptides and present them to T lymphocytes. MHC class I molecules consist of two polypeptide chains, an a or heavy chain, and β-2-Microglobulin, a non-covalently associated protein. Cytotoxic T lymphocytes bind antigenic peptides presented by MHC class I molecules. Antigens that bind to MHC class I molecules are typically 8-10 residues in length and are stabilized in a peptide binding groove. MHC class II molecules are encoded by polymorphic MHC genes and consist of a non-covalent complex of an a and b chain. Helper T lymphocytes bind antigenic peptides presented by MHC class II molecules. MHC class II molecules bind 13-18 amino acid antigenic peptides. Accumulating in endosomal/lysosomal compartments and on the surface of B cells, HLA-DM and -DO molecules regulate binding of exogenous peptides to class II molecules (HLA-DR) by sustaining a conformation that favors peptide exchange. The differential structural properties of MHC class I and class II molecules account for their respective roles in activating different populations of T lymphocytes.
Background References
1. Maglietta A et al. The Immune Landscapes of Polypoid and Nonpolypoid Precancerous Colorectal Lesions. PLoS One 11:e0159373 (2016).
2. Placke T et al. Platelet-derived MHC class I confers a pseudonormal phenotype to cancer cells that subverts the antitumor reactivity of natural killer immune cells. Cancer Res 72:440-8 (2012).
Subcellular Location
Cell membrane.
Synonyms
A 28 antibody
A 9 antibody
Antigen presenting molecule antibody
Aw 24 antibody
Aw 68 antibody
CLASS I HISTOCOMPATIBILITY ANTIGEN antibody
H2 K1 antibody
H2K antibody
HLA A antibody
HLA class I histocompatibility antigen A 1 alpha chain antibody
ExpandImages
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☑ Relative expression (RE)
Western blot analysis of MHC class I on different lysates with Rabbit anti-MHC class I antibody (HA750347) at 1/1,000 dilution.
Lane 1: A431 cell lysate
Lane 2: HL-60 cell lysate
Lane 3: K-562 cell lysate (low expression)
Lane 4: Jurkat cell lysate
Lane 5: Raji cell lysate
Lane 6: THP-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 41 kDa
Observed band size: 45 kDa
Exposure time: 1 minute; 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 (HA750347) at 1/1,000 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. -
☑ Knockdown (KD)
Western blot analysis of MHC class I on different lysates with Rabbit anti-MHC class I antibody (HA750347) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-MHC class I KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 41 kDa
Observed band size: 41 kDa
Exposure time: 30 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 (HA750347) 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. -
☑ Relative expression (RE)
Immunocytochemistry analysis of HL-60 cells and K-562 (low expression) labeling MHC class I with Rabbit anti-MHC class I antibody (HA750347) 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-MHC class I antibody (HA750347) 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. -
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-MHC class I antibody (HA750347) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750347) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-MHC class I antibody (HA750347) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750347) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-MHC class I antibody (HA750347) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750347) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-MHC class I antibody (HA750347) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750347) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling MHC class I with Rabbit anti-MHC class I antibody (HA750347) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750347, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). -
Flow cytometric analysis of HL-60 cells labeling MHC class I.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750347, 1μg/mL) (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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"