HLA-E Monoclonal antibody

• 

  WB analysis of THP-1 using 66530-1-Ig

  THP-1, Ramos, Daudi, and Raji cells were subjected to SDS PAGE followed by western blot with 66530-1-Ig (HLA-E antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat using 66530-1-Ig

  Jurkat cells were subjected to SDS PAGE followed by western blot with 66530-1-Ig (HLA-E antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HUVEC using 66530-1-Ig

  HUVEC cells were subjected to SDS PAGE followed by western blot with 66530-1-Ig (HLA-E antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human placenta using 66530-1-Ig

  human placenta tissue were subjected to SDS PAGE followed by western blot with 66530-1-Ig (HLA-E antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of THP-1 using 66530-1-Ig

  IP result of anti-HLA-E (IP:66530-1-Ig, 4ug; Detection:66530-1-Ig 1:3000) with THP-1 cells lysate 1720 ug.

• 

  IHC staining of human tonsillitis using 66530-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66530-1-Ig (HLA-E antibody) at dilution of 1:400 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 66530-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66530-1-Ig (HLA-E antibody) at dilution of 1:400 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human placenta using 66530-1-Ig

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 66530-1-Ig (HLA-E antibody) at dilution of 1:400 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human placenta using 66530-1-Ig

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 66530-1-Ig (HLA-E antibody) at dilution of 1:400 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of human tonsillitis using 66530-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using HLA-E antibody (66530-1-Ig, Clone: 1A4G3 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

HLA-E Monoclonal Antibody for WB, IHC, IF-P, IP, ELISA

Cat No. 66530-1-Ig

Clone No.1A4G3

Publications(3)

Host / Isotype

Mouse / IgG2a

Reactivity

human

Applications

WB, IHC, IF-P, IP, ELISA

Synonyms

1A4G3, HLA 6.2, HLA E, HLA-6.2, HLAE

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Tested Applications

• Positive WB detected in: THP-1 cells, Jurkat cells, HUVEC cells, human placenta tissue, Ramos cells, Daudi cells, Raji cells
• Positive IP detected in: THP-1 cells
• Positive IHC detected in: human tonsillitis tissue, human placenta tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: human tonsillitis tissue

Recommended dilution

• Western Blot (WB): WB : 1:2500-1:10000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 1 publications below
• IHC: See 1 publications below
• IF: See 1 publications below

Product Information

66530-1-Ig targets HLA-E in WB, IHC, IF-P, IP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag6724

  Product name: Recombinant human HLA-E protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 23-303 aa of BC002578

  Sequence: SHSLKYFHTSVSRPGRGEPRFISVGYVDDTQFVRFDNDAASPRMVPRAPWMEQEGSEYWDRETRSARDTAQIFRVNLRTLRGYYNQSEAGSHTLQWMHGCELGPDRRFLRGYEQFAYDGKDYLTLNEDLRSWTAVDTAAQISEQKSNDASEAEHQRAYLEDTCVEWLHKYLEKGKETLLHLEPPKTHVTHHPISDHEATLRCWALGFYPAEITLTWQQDGEGHTQDTELVETRPAGDGTFQKWAAVVVPSGEEQRYTCHVQHEGLPEPVTLRWKPASQPTI

• Full Name: major histocompatibility complex, class I, E
• Calculated Molecular Weight: 40 kDa
• Observed Molecular Weight: 40 kDa
• GenBank Accession Number: BC002578
• Gene Symbol: HLA-E
• Gene ID (NCBI): 3133
• RRID: AB_2881893
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P13747
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Human major histocompatibility complex (MHC) antigens, also referred to as human leukocyte antigens (HLA), are encoded by genes located on the short arm of chromosome 6 (6p21.3). There are two classes of HLA antigens: class I and class II. This class I molecules are membrane glycoproteins composed of a heavy (alpha) chain which is encoded by a HLA class I gene, and β2-microglobulin light (beta) chain. The most extensively characterized members of the HLA class I gene family are the genes encoding the major transplantation antigenes, HLA-A, B and C. HLA-E is a non-classical MHC class I molecule. HLA-E is frequently overexpressed in tumor diseases, transplants and virus-infected cells and represents an immunomodulatory molecule by binding to the receptors CD94/NKG2A, -B and -C on NK and T cells. Due to its immune suppressive features HLA-E expression might represent an important mechanism of tumors to escape immune surveillance.(PMID: 667938; 3375250; 2249951; 27589686)

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	IF	Cell; Single-cell dissection of the human motor and prefrontal cortices in ALS and FTLD; Authors - S Sebastian Pineda
human	IHC	Nat Med; Allogeneic NK cells with a bispecific innate cell engager in refractory relapsed lymphoma: a phase 1 trial; Authors - Yago Nieto
human	WB	Cancer Cell; Immune checkpoint HLA-E:CD94-NKG2A mediates evasion of circulating tumor cells from NK cell surveillance; Authors - Xiaowei Liu ; KD Validated