MLH1 Recombinant Rabbit Monoclonal Antibody [PD01-61]
Catalog# HA721325
MLH1 Recombinant Rabbit Monoclonal Antibody [PD01-61]
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WB
-
IHC-P
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Human
-
Mouse
-
Rat
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unconjugated
製品概要
製品名
MLH1 Recombinant Rabbit Monoclonal Antibody [PD01-61]
抗体のタイプ
Recombinant Rabbit monoclonal Antibody
免疫原
Synthetic peptide corresponding to Human MLH1 aa 700-756.
交差反応性（対応種属）
Human, Mouse, Rat
検証済みアプリケーション
WB, IHC-P
標的分子量
Predicted band size: 85 kDa
ポジティブコントロール
HeLa cell lysate, HCT 116 cell lysate, A549 cell lysate, C2C12 cell lysate, NIH/3T3 cell lysate, human appendix tissue, human colon tissue, human B-cell lymphoblastic lymphoma tissue, human NKT cell lymphoma tissue, mouse testis tissue, mouse thymus tissue, human colon carcinoma tissue, rat small intestine tissue.
コンジュゲーション
unconjugated
クローン番号
PD01-61
RRID
製品の特徴
形態
Liquid
濃度
保存方法
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
保存バッファー
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
アイソタイプ
IgG
精製方法
Protein A affinity purified.
推奨希釈倍率
-
WB
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1:2,000
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IHC-P
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1:200-1:1,000
ターゲット
機能
DNA-mismatch repair (MMR) is an essential process in maintaining genetic stability. Lack of a functional DNA-mismatch repair pathway is a common characteristic of several different types of human cancers, either due to an MMR gene mutation or promoter methylation gene silencing. MLH1 is an integral part of the protein complex responsible for mismatch repair and is expressed in lymphocytes, heart, colon, breast, lung, spleen, testis, prostate, thyroid and gall bladder tissues, and is methylated in several ovarian tumors. Loss of MLH1 protein expression is associated with a mutated phenotype, microsatellite instability and a predisposition to cancer. In hereditary nonpolyposis colorectal cancer (HNPCC), an autosomal dominant inherited cancer syndrome that signifies a high risk of colorectal and various other types of cancer, the MLH1 gene exhibits a pathogenic mutation. Certain cancer cell lines, including leukemia CCRF-CEM, colon HCT 116 and KM12, and ovarian cancers SK-OV-3 and IGROV-1, show complete deficiency of MLH1, while MLH1 is expressed in 60% of melanomas, 70% of noninvasive squamous cell carcinomas and 30% of invasive squamous cell carcinomas.
背景・参考文献
1. Guan J et al. MLH1 Deficiency-Triggered DNA Hyperexcision by Exonuclease 1 Activates the cGAS-STING Pathway. Cancer Cell. 2021 Jan
2. Cannavo E et al. Regulation of the MLH1-MLH3 endonuclease in meiosis. Nature. 2020 Oct
配列相同性
Belongs to the DNA mismatch repair MutL/HexB family.
組織特異性
Colon, lymphocytes, breast, lung, spleen, testis, prostate, thyroid, gall bladder and heart.
サブセルラー局在
Nucleus.
別名
COCA 2 antibody
COCA2 antibody
DNA mismatch repair protein Mlh1 antibody
FCC 2 antibody
FCC2 antibody
hMLH 1 antibody
hMLH1 antibody
HNPCC 2 antibody
HNPCC antibody
HNPCC2 antibody
詳細を見る画像
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☑ Relative expression (RE)
Western blot analysis of MLH1 on different lysates with Rabbit anti-MLH1 antibody (HA721325) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HCT 116 cell lysate (negative)
Lane 3: A549 cell lysate
Lysates/proteins at 15 µg/Lane.
Predicted band size: 85 kDa
Observed band size: 85 kDa
Exposure time: 2 minutes; 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 (HA721325) at 1/2,000 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. -
Western blot analysis of MLH1 on different lysates with Rabbit anti-MLH1 antibody (HA721325) at 1/2,000 dilution.
Lane 1: C2C12 cell lysate
Lane 2: NIH/3T3 cell lysate
Lysates/proteins at 30 µg/Lane.
Predicted band size: 85 kDa
Observed band size: 85 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 (HA721325) at 1/2,000 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 MLH1 on different lysates with Rabbit anti-MLH1 antibody (HA721325) at 1/2,000 dilution.
Lane 1: Hela-si NT cell lysate (10 µg/Lane)
Lane 2: Hela-si MLH1 cell lysate (10 µg/Lane)
Predicted band size: 85 kDa
Observed band size: 85 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
HA721325 was shown to specifically react with MLH1 in Hela-si NT cells. Weakened band was observed when Hela-si MLH1 sample was tested. Hela-si NT and Hela-si MLH1 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (HA721325, 1/2,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were 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. -
Immunohistochemical analysis of paraffin-embedded human appendix tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 colon tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 B-cell lymphoblastic lymphoma tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 NKT cell lymphoma tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 mouse testis tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 mouse thymus tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 colon carcinoma tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 rat small intestine tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 colon cancer tissue using anti-MLH1 antibody (1/100) performed on the Dako Omnis.
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