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HLA-ABC (MHC I) Recombinant Rabbit Monoclonal Antibody [PSH12-02] - BSA and Azide free

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Specification

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Catalog# HA751435

HLA-ABC (MHC I) Recombinant Rabbit Monoclonal Antibody [PSH12-02] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

HLA-ABC (MHC I) Recombinant Rabbit Monoclonal Antibody [PSH12-02] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human HLA-B aa 110-309.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 40 kDa

Positive Control

Raji cell lysate, THP-1 cell lysate, Jurkat cell lysate, HL-60 cell lysate, HepG2 cell lysate, Ramos cell lysate, Mouse spleen tissue lysate, Mouse lung tissue lysate, Rat spleen tissue lysate, Rat lung tissue lysate, Jurkat, Raji, human spleen tissue, human stomach tissue, mouse spleen tissue, mouse stomach tissue, rat spleen tissue, rat stomach tissue.

Conjugation

unconjugated

Clone Number

PSH12-02

Product Features

Form

Liquid

Concentration

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

  • 1:5,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:2,000

Target

Function

MHC class I molecules are one of two primary classes of major histocompatibility complex (MHC) molecules (the other being MHC class II) and are found on the cell surface of all nucleated cells in the bodies of vertebrates. They also occur on platelets, but not on red blood cells. Their function is to display peptide fragments of proteins from within the cell to cytotoxic T cells; this will trigger an immediate response from the immune system against a particular non-self antigen displayed with the help of an MHC class I protein. Because MHC class I molecules present peptides derived from cytosolic proteins, the pathway of MHC class I presentation is often called cytosolic or endogenous pathway. In humans, the HLAs corresponding to MHC class I are HLA-A, HLA-B, and HLA-C.

Background References

1. Singh S et al. Downregulation of HLA-ABC expression through promoter hypermethylation and downmodulation of MIC-A/B surface expression in LMP2A-positive epithelial carcinoma cell lines. Sci Rep. 2020 Mar

Subcellular Location

Cell membrane, Endoplasmic reticulum membrane.

UNIPROT

SwissProt: P04439 Human

SwissProt: P01889 Human

SwissProt: P10321 Human

Synonyms

Antigen Presenting Molecule antibody

Cw*1701 antibody

D6S204 antibody

FLJ27082 antibody

HLA A antibody

HLA B antibody

HLA C antibody

HLA class 1 A antibody

HLA class 1 B antibody

HLA class 1 C antibody

Expand

Images

  • Western blot analysis of HLA-ABC (MHC I) on different lysates with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/5,000 dilution.<br /><br />Lane 1: Raji cell lysate (20 µg/Lane)<br />Lane 2: THP-1 cell lysate (20 µg/Lane)<br />Lane 3: Jurkat cell lysate (20 µg/Lane)<br />Lane 4: HL-60 cell lysate (20 µg/Lane)<br />Lane 5: HepG2 cell lysate (20 µg/Lane)<br />Lane 6: Ramos cell lysate (20 µg/Lane)<br />Lane 7: Mouse spleen tissue lysate (40 µg/Lane)<br />Lane 8: Mouse lung tissue lysate (40 µg/Lane)<br />Lane 9: Rat spleen tissue lysate (40 µg/Lane)<br />Lane 10: Rat lung tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 40 kDa<br />Observed band size: 45 kDa<br /><br />Exposure time: 10 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/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/5,000 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 HLA-ABC (MHC I) on different lysates with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/5,000 dilution.

    Lane 1: Raji cell lysate (20 µg/Lane)
    Lane 2: THP-1 cell lysate (20 µg/Lane)
    Lane 3: Jurkat cell lysate (20 µg/Lane)
    Lane 4: HL-60 cell lysate (20 µg/Lane)
    Lane 5: HepG2 cell lysate (20 µg/Lane)
    Lane 6: Ramos cell lysate (20 µg/Lane)
    Lane 7: Mouse spleen tissue lysate (40 µg/Lane)
    Lane 8: Mouse lung tissue lysate (40 µg/Lane)
    Lane 9: Rat spleen tissue lysate (40 µg/Lane)
    Lane 10: Rat lung tissue lysate (40 µg/Lane)

    Predicted band size: 40 kDa
    Observed band size: 45 kDa

    Exposure time: 10 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 (HA751435) at 1/5,000 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 HLA-ABC (MHC I) with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 HLA-ABC (MHC I) with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-HLA-ABC (MHC I) antibody (HA751435) 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 (HA601187, 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.

  • Immunocytochemistry analysis of Raji cells labeling HLA-ABC (MHC I) with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 Raji cells labeling HLA-ABC (MHC I) with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-HLA-ABC (MHC I) antibody (HA751435) 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 (HA601187, 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 spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 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 spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/2,000 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 (HA751435) at 1/2,000 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 stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 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 stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/2,000 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 (HA751435) at 1/2,000 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 mouse spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 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 mouse spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/2,000 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 (HA751435) at 1/2,000 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 mouse stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 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 mouse stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/2,000 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 (HA751435) at 1/2,000 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 rat spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 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 rat spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/2,000 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 (HA751435) at 1/2,000 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 rat stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751435" style="font-weight: bold;text-decoration: underline;">HA751435</a>) at 1/2,000 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 rat stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA751435) at 1/2,000 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 (HA751435) at 1/2,000 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.

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