MHC Class I Antibody (OX18) - BSA Free

Novus Biologicals | Catalog # NB120-6405

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Rat

Cited:

Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, ELISA, Block/Neutralize, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Electron Microscopy, CyTOF-ready

Cited:

Immunohistochemistry-Frozen, Block/Neutralize, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # OX18

Format

BSA Free
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Product Specifications

Immunogen

Rat spleen glycoproteins

Localization

Cell Membrane; Type I membrane protein

Specificity

Recognizes a monomorphic determinant of rat MHC Class I (RT1A), expressed by all rat strains. However, quantitative measurements suggest that not all of the class I molecules are recognised.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for MHC Class I Antibody (OX18) - BSA Free

Immunocytochemistry/ Immunofluorescence: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Immunocytochemistry/ Immunofluorescence: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

MHC-Class-I-Antibody-OX18-Immunocytochemistry-Immunofluorescence-NB120-6405-img0009.jpg
Immunohistochemistry-Paraffin: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Immunohistochemistry-Paraffin: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Immunohistochemistry-Paraffin: MHC Class I Antibody (OX18) [NB120-6405] - Analysis of FFPE rat brain cerebellum using MHC Class I (OK18) antibody at 1:200 on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Endothelial staining was observed. Staining was performed by Histowiz.
Flow Cytometry: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Flow Cytometry: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Flow Cytometry: MHC Class I Antibody (OX18) [NB120-6405] - Analysis using the FITC conjugate of NB120-6405. Staining of rat spleen cells with mouse anti-rat RT1-A (OX18).
Immunocytochemistry/ Immunofluorescence: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Immunocytochemistry/ Immunofluorescence: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Immunocytochemistry/Immunofluorescence: MHC Class I Antibody (OX18) [NB120-6405] - PC-12 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-MHC Class I (OX18) NB120-6405 at a 1:100 dilution overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunocytochemistry/ Immunofluorescence: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Immunocytochemistry/ Immunofluorescence: MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Immunocytochemistry/Immunofluorescence: MHC Class I Antibody (OX18) [NB120-6405] - PC12 cells were fixed for 10 minutes using 4% PFA and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-MHC Class I Antibody (OX18) at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Flow (Intracellular): MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Flow (Intracellular): MHC Class I Antibody (OX18) - BSA Free [NB120-6405]

Flow (Intracellular): MHC Class I Antibody (OX18) [NB120-6405] - PC-12 cells were stained with MHC Class I NB120-6405 (Blue) and a matched isotype control NBP2-27287 (Orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by DyLight488-conjugated anti-mouse secondary antibody.
MHC Class I Antibody (OX18) - BSA Free

Immunohistochemistry: MHC Class I Antibody (OX18) - BSA Free [NB120-6405] -

Effects of anodal tDCS on protein expression. Representative immunohistochemical images of Ox18 + cells in the SVZ ipsilateral to anodal tDCS or sham stimulation. Staining for Ox18 (MHC I) in the ipsilateral SVZ revealed more Ox18 + cells in animals treated by anodal tDCS (left panel) compared to sham stimulation (right panel) by trend (scale bars = 100 μm). Results are displayed as mean ± SEM.
MHC Class I Antibody (OX18) - BSA Free

Immunohistochemistry: MHC Class I Antibody (OX18) - BSA Free [NB120-6405] -

Immunohistochemistry: MHC Class I Antibody (OX18) - BSA Free [NB120-6405] - Effects of anodal tDCS on protein expression. Representative immunohistochemical images of Ox18 + cells in the SVZ ipsilateral to anodal tDCS or sham stimulation. Staining for Ox18 (MHC I) in the ipsilateral SVZ revealed more Ox18 + cells in animals treated by anodal tDCS (left panel) compared to sham stimulation (right panel) by trend (scale bars = 100 μm). Results are displayed as mean ± SEM. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31708742), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
MHC Class I Antibody (OX18) - BSA Free

Immunocytochemistry/ Immunofluorescence: MHC Class I Antibody (OX18) - BSA Free [NB120-6405] -

Major histocompatibility complex (MHC) I and II as well as Transporter associated with antigen presentation II (TAPII) were analyzed, using immunocytochemistry on rat Schwann cells (SCs). Corresponding merges are shown in the bottom rows. Treatment of SCs with IL-17 was performed at concentrations of 0.5 and 50 ng/mL. Graphs to the right show densitometry quantification. SCs showed expression of MHCI > TAPII > MHCII, which increased after IL-17 treatment. (A) MHCI was mainly detected in the cytoplasm and the expression increased in a dose-dependent manner after IL-17 treatment, significant for 0.5 ng/mL and 50 ng/mL (**P ≤0.01). (B) MHCII revealed a fainter basic expression emphasizing the nucleus and was found significantly increased after 0.5 ng/mL IL-17 stimulation (**P ≤ 0.01). (C) TAPII was detected in the nucleus and cytoplasm. We detected a dose-dependent tendency but no significantly increased expression after IL-17 stimulation. For MHCI, TAPII and MHCII and the concentrations applied, analysis of variance between the independent experiments revealed no significant difference. DAPI, 4′, 6-diamidino-2-phenylindole. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24678820), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for MHC Class I Antibody (OX18) - BSA Free

Application
Recommended Usage

Block/Neutralize

reported in scientific literature (PMID 1698855)

ELISA

1:100-1:2000. Use reported in scientific literature (PMID 2783579)

Electron Microscopy

reported in scientific literature (PMID 3044648)

Flow Cytometry

1:50-1:100

Immunocytochemistry/ Immunofluorescence

1:10-1:500. Use reported in scientific literature (PMID 24678820)

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

1:10-1:500
Application Notes
The epitope recognized by this anti-rat RT1-A antibody may be sensitive to formaldehyde fixation and tissue processing. If possible, the use of acetone fixation for frozen sections is recommended. This antibody is CyTOF ready.

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Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: MHC Class I

MHC Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. MHC class I antigens are heterodimers consisting of one alpha chain (44kDa) with beta 2 microglobulin (11.5 kDa). The antigen is expressed by all somatic cells at varying levels. MHC Class I molecules are expressed on most nucleated cells where they present endogenously synthesized antigenic peptides to CD8+ T lymphocytes, which are usually cytotoxic T cells. Fibroblasts or neurons however only show a low level of antigen.

Alternate Names

DKFZp686P19218, EA1.2, EA2.1, HLA class I histocompatibility antigen, alpha chain E, HLA class I histocompatibility antigen, E alpha chain, HLA-6.2, HLAE, lymphocyte antigen, major histocompatibility complex, class I, E, MHC, MHC class I antigen E, MHC HLA-E alpha-1, MHC HLA-E alpha-2.1, QA1, Clone OX18, MHC Class I RT1-A, OX18, RT1-A, RT1-A (OX18)

Gene Symbol

HLA-E

Additional MHC Class I Products

Product Documents for MHC Class I Antibody (OX18) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for MHC Class I Antibody (OX18) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for MHC Class I Antibody (OX18) - BSA Free

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Protocols

View specific protocols for MHC Class I Antibody (OX18) - BSA Free (NB120-6405):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

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