HLA class I ABC Monoclonal antibody

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  WB analysis using 66013-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

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  WB analysis of U-937 using 66013-1-Ig

  U-937 cells were subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of Raji using 66013-1-Ig

  Raji cells were subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 66013-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 66013-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human white blood using 66013-1-Ig

  human white blood cells were subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HepG2 using 66013-1-Ig

  HepG2 cells were subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of pig spleen using 66013-1-Ig

  pig spleen tissue was subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of NCCIT using 66013-1-Ig

  NCCIT cells were subjected to SDS PAGE followed by western blot with 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of HepG2 using 66013-1-Ig

  IP result of anti-HLA class I ABC (IP:66013-1-Ig, 4ug; Detection:66013-1-Ig 1:4000) with HepG2 cells lysate 2000 ug.

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  IHC staining of human tonsillitis using 66013-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 66013-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 66013-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 66013-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human stomach cancer using 66013-1-Ig

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human stomach cancer using 66013-1-Ig

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 66013-1-Ig (HLA class I ABC antibody) at dilution of 1:10000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of Raji using 66013-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed Raji cells using HLA class I ABC antibody (66013-1-Ig, Clone: 5C5B7 ) at dilution of 1:800 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  FC experiment of Raji using 66013-1-Ig

  1X10^6 Raji cells were surface stained with 0.4 ug Anti-Human HLA class I ABC (66013-1-Ig, Clone:5C5B7) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2b Isotype Control (MPC-11) (65128-1-Ig, Clone: MPC-11) (blue). Cells were not fixed.

• Featured Product
• KD/KO Validated

HLA class I ABC Monoclonal Antibody for WB, IHC, IF/ICC, FC, IP, ELISA

Cat No. 66013-1-Ig

Clone No.5C5B7

Publications(16)

Host / Isotype

Mouse / IgG2b

Reactivity

human, pig

Applications

WB, IHC, IF/ICC, FC, IP, ELISA

Synonyms

HLA-A, 5C5B7, HLA A, HLA class I, HLA class I (HLA-A)

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Tested Applications

• Positive WB detected in: HeLa cells, HepG2 cells, human white blood, NCCIT cells, pig spleen tissue, U-937 cells, Raji cells, HEK-293 cells, Jurkat cells
• Positive IP detected in: HepG2 cells
• Positive IHC detected in: human tonsillitis tissue, human liver cancer tissue, human stomach cancer 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/ICC detected in: Raji cells
• Positive FC detected in: Raji cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:1000-1:10000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC): FC : 0.40 ug per 10^6 cells in a 100 µl suspension

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 8 publications below
• IHC: See 5 publications below
• IF: See 3 publications below
• IP: See 1 publications below

Product Information

66013-1-Ig targets HLA class I ABC in WB, IHC, IF/ICC, FC, IP, ELISA applications and shows reactivity with human, pig samples.

• Tested Reactivity: human, pig
• Cited Reactivity: human, pig
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag7488

  Product name: Recombinant human HLA class I (HLA-A) protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 28-303 aa of BC003069

  Sequence: SMRYFFTSVSRPGRGEPRFIAVGYVDDTQFVRFDSDAASQKMEPRAPWIEQEGPEYWDQETRNMKAHSQTDRANLGTLRGYYNQSEDGSHTIQIMYGCDVGPDGRFLRGYRQDAYDGKDYIALNEDLRSWTAADMAAQITKRKWEAVHAAEQRRVYLEGRCVDGLRRYLENGKETLQRTDPPKTHMTHHPISDHEATLRCWALGFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVVVPSGEEQRYTCHVQHEGLPKPLTLRWELSSQ

• Full Name: major histocompatibility complex, class I, A
• Calculated Molecular Weight: 41 kDa
• Observed Molecular Weight: 40-44 kDa
• GenBank Accession Number: BC003069
• Gene Symbol: HLA-A
• Gene ID (NCBI): 3105
• RRID: AB_11042593
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P04439
• 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 (HLA-A, B and C) and class II (HLA-D). This class I molecules are polymorphic membrane glycoproteins composed of a heavy (alpha) chain (44 kDa) which is encoded by a HLA class I gene (HLA-A, B or C), and β2-microglobulin light (beta) chain (12 kDa). They are involved in the presentation of foreign antigens to the immune system. (PMID: 667938; 3375250)

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	IF	Front Med; Distinct immune escape and microenvironment between RG-like and pri-OPC-like glioma revealed by single-cell RNA-seq analysis; Authors - Weiwei Xian
		Oncotarget; Combination epidermal growth factor receptor variant III peptide-pulsed dendritic cell vaccine with miR-326 results in enhanced killing on EGFRvIII-positive cells.; Authors - Jianlong Li
human	WB	J Tissue Eng Regen Med; Sequential hydrophile and lipophile solubilization as an efficient method for decellularization of porcine aortic valve leaflets: structure, mechanical property and biocompatibility study.; Authors - Wei-Hua Qiao
human	WB	J Biol Chem; The transferrin receptor CD71 regulates type II CD38, revealing tight topological compartmentalization of intracellular cyclic ADP-ribose production.; Authors - Qi Wen Deng ; KD Validated
pig	IHC	Tissue Eng Part A; DECELLULARIZATION OF MASSIVE BONE ALLOGRAFTS BY PERFUSION: A NEW PROTOCOL FOR TISSUE ENGINEERING; Authors - Robin Evrard
human	IHC	Asian Pac J Cancer Prev; Loss of heterozygosity at 6p21 and HLA class I expression in esophageal squamous cell carcinomas in China.; Authors - Zhao Xiaogang X