HA601135

MLH1 Mouse Monoclonal Antibody [A9F7]

cat.: HA601135

Product Type:	Mouse monoclonal IgG2b, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P
Clonality:	Monoclonal
Clone number:	A9F7

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 85 kDa
Isotype:	IgG2b
Immunogen:	Recombinant protein within human MLH1 aa 452-751 / 751.
Positive control:	HeLa cell lysate, A549 cell lysate, HepG2 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, F9 cell lysate, RAW264.7 cell lysate, C6 cell lysate, A431 cell lysate, Daudi cell lysate, K-562 cell lysate, HL-60 cell lysate, SW480 cell lysate, human colon carcinoma tissue, mouse colon tissue, rat small intestine tissue.
Subcellular location:	Nucleus, Chromosome.
Recommended Dilutions: WB IHC-P	1:1,000-1:2,000 1:1,000
Uniprot #:	SwissProt: P40692 Human \| Q9JK91 Mouse \| P97679 Rat
Alternative names:	COCA 2 COCA2 DNA mismatch repair protein Mlh1 FCC 2 FCC2 hMLH 1 hMLH1 HNPCC 2 HNPCC HNPCC2 MGC5172 MLH 1 MLH1 MLH1_HUMAN MutL homolog 1 (E. coli) MutL homolog 1 MutL homolog 1 colon cancer nonpolyposis type 2 MutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli) MutL protein homolog 1 MutL, E. coli, homolog of, 1

Images

Fig1: Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HCT 116 cell lysate (negative) (15 µg/Lane)
Lane 3: A549 cell lysate (15 µg/Lane)
Lane 4: HepG2 cell lysate (15 µg/Lane)
Lane 5: Jurkat cell lysate (15 µg/Lane)

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: Lane 1-5 (left): 24 seconds; Lane 1-5 (right): 2 minutes;

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 (HA601135) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: F9 cell lysate (20 µg/Lane)
Lane 3: RAW264.7 cell lysate (20 µg/Lane)
Lane 4: C6 cell lysate (20 µg/Lane)

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: 3 minutes 10 seconds;

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 (HA601135) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.

Fig3: Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

Lane 1: A431 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: Daudi cell lysate (20 µg/Lane)
Lane 4: K-562 cell lysate (20 µg/Lane)
Lane 5: HL-60 cell lysate (20 µg/Lane)
Lane 6: SW480 cell lysate (20 µg/Lane)
Lane 7: HCT 116 cell lysate (Negative) (20 µg/Lane)

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: 20 seconds;

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 (HA601135) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.

Fig4: Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/2,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si MLH1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: 50 seconds;

4-20% SDS-PAGE gel.

HA601135 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 (HA601135, 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-mouse IgG-HRP Secondary Antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.

	Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA601135) 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA601135) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA601135) 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.