Name: Anti-MSH6 antibody [EPR3945]
SKU: ab92471
Rating: 5 (5 reviews)

Anti-MSH6 antibody [EPR3945] is a rabbit recombinant monoclonal antibody that is used to detect MSH6 in ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse, Rat samples.

- Recombinant format ensures unrivaled batch-to-batch consistency
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity and sensitivity confirmed in immunohistochemistry (IHC) with multi-tissue microarray (TMA) validation
- Cited in over 55 publications

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Alexa Fluor® 594 Anti-MSH6 antibody [EPR3945]
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Alexa Fluor® 488 Anti-MSH6 antibody [EPR3945]
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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] (AB92471), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	WB	ICC/IF
Human	Tested	Tested	Tested
Mouse	Expected	Expected	Expected
Rat	Tested	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/500	Notes For unpurified, use 1/100 - 1/250. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Human	Dilution info 1/500	Notes For unpurified, use 1/100 - 1/250. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes For unpurified, use 1/100 - 1/250. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/1000 - 1/10000	Notes -
Species Human	Dilution info 1/1000 - 1/10000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/10000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1 µg/mL	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -
Species Mouse	Dilution info 1 µg/mL	Notes -

Associated Products

Select an associated product type

6 products for Alternative Product

Target data

See full target information MSH6

Function

Component of the post-replicative DNA mismatch repair system (MMR). Heterodimerizes with MSH2 to form MutS alpha, which binds to DNA mismatches thereby initiating DNA repair. When bound, MutS alpha bends the DNA helix and shields approximately 20 base pairs, and recognizes single base mismatches and dinucleotide insertion-deletion loops (IDL) in the DNA. After mismatch binding, forms a ternary complex with the MutL alpha heterodimer, which is thought to be responsible for directing the downstream MMR events, including strand discrimination, excision, and resynthesis. ATP binding and hydrolysis play a pivotal role in mismatch repair functions. The ATPase activity associated with MutS alpha regulates binding similar to a molecular switch: mismatched DNA provokes ADP-->ATP exchange, resulting in a discernible conformational transition that converts MutS alpha into a sliding clamp capable of hydrolysis-independent diffusion along the DNA backbone. This transition is crucial for mismatch repair. MutS alpha may also play a role in DNA homologous recombination repair. Recruited on chromatin in G1 and early S phase via its PWWP domain that specifically binds trimethylated 'Lys-36' of histone H3 (H3K36me3): early recruitment to chromatin to be replicated allowing a quick identification of mismatch repair to initiate the DNA mismatch repair reaction.

Alternative names

GTBP, MSH6, DNA mismatch repair protein Msh6, hMSH6, G/T mismatch-binding protein, MutS protein homolog 6, MutS-alpha 160 kDa subunit, GTMBP, p160

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR3945
Purification technique: Affinity purification Protein A
Dissociation constant: 2.3 x 10^-9 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

To see more of the key markers and tools you need to study the hallmarks of cancer, including genome instability and mutation, please visit the following page.

Product Specifications
Anti-MSH6 antibody [EPR3945] (ab92471) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ICC/IF, IHC-P, WB in human, mouse, rat samples.
Anti-MSH6 antibody [EPR3945] (ab92471) specifically detects MSH6 (UniProt ID: P52701; Molecular weight: 153kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-MSH6 antibody [EPR3945] (ab92471) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-MSH6 antibody [EPR3945] (ab92471) has been confirmed by testing in knockout samples.
Anti-MSH6 antibody [EPR3945] (ab92471) has been cited over 58 times in peer reviewed journals and is trusted by the scientific community.
Anti-MSH6 antibody [EPR3945] (ab92471) has 5 independent reviews from customers.

Related Products
Antibody clone EPR3945 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 647, Alexa Fluor® 488, Alexa Fluor® 594, Alexa Fluor® 555, Alexa Fluor® 750 (ab198334, ab311007, ab311771, ab313252, ab321758).

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

19 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] (ab92471)
Immunohistochemical staining of paraffin embedded human colon with purified ab92471 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
Lanes 1- 2: Merged signal (red and green). Green - ab92471 observed at 160 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
ab92471 was shown to react with MSH6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human MSH6 knockout HeLa cell line ab255410 (knockout cell lysate Human MSH6 knockout HeLa cell lysate ab263763) was used. Wild-type HeLa and MSH6 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab92471 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: MSH6 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human MSH6 knockout HeLa cell line (Human MSH6 knockout HeLa cell line ab255410)
Performed under reducing conditions.
Predicted band size: 153 kDa
Observed band size: 160 kDa
Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR3945] (ab92471)
ab92471 staining MSH6 in wild-type HAP1 cells (top panel) and MSH6 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92471 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
Lanes 1 - 4: Merged signal (red and green). Green - ab92471 observed at 160 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab92471 was shown to specifically react with MSH6 in wild-type HAP1 cells. No band was observed when MSH6 knockout samples were used. Wild-type and MSH6 knockout samples were subjected to SDS-PAGE. ab92471 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution
Lane 1: Wild-type HAP1 cell lysate at 20 µg
Lane 2: MSH6 knockout HAP1 cell lysate at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: A431 cell lysate at 20 µg
Predicted band size: 153 kDa
Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
Different batches of ab92471 were tested on Rat brain lysate at 0.2 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 160 kDa.
All lanes: Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
Predicted band size: 153 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] (ab92471)
Immunohistochemical staining of paraffin embedded rat liver with purified ab92471 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution
All lanes: Rat brain at 10 µg
Secondary
All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 153 kDa
Observed band size: 160 kDa
This image is courtesy of a customer review by Serena Bologna.
Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
All lanes: Western blot - Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution
Lane 1: HAP1 WT cell lysate at 50 µg
Lane 2: MSH6 KO HAP1 cell lysate at 50 µg
Secondary
All lanes: donkey anti-rabbit IgG-HRP at 1/3000 dilution
Developed using the ECL technique.
Predicted band size: 153 kDa
Exposure time: 5min
Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: purified at 1/6000 dilution
All lanes: SW480 cell lysate at 10 µg
Secondary
All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 153 kDa
Observed band size: 160 kDa
Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR3945] (ab92471)
ab92471 staining MSH6 in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92471 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-MSH6 antibody [EPR3945] (ab92471) at 1/2000 dilution
All lanes: SW480 cell lysate at 10 µg
Secondary
All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 153 kDa
Observed band size: 160 kDa
Image courtesy of a customer review submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR3945] (ab92471)
Unpurified ab92471 (1/500) staining MSH6 in asynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please see abreview.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] (ab92471)
Unpurified ab92471, at a 1/100 dilution, detecting MSH6 in paraffin embedded Human colonic adenocarcinoma tissue by immunohistochemistry. Detection used HRP conjugated anti rabbit antibody.
Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Western blot - Anti-MSH6 antibody [EPR3945] (ab92471)
Secondary antibody - goat anti-rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab6721)
All lanes: Western blot - Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution
Lane 1: A431 cell lysate at 10 µg
Lane 2: HeLa cell lysate at 10 µg
Lane 3: SW480 cell lysate at 10 µg
Secondary
All lanes: HRP labelled goat anti-rabbit antibody at 1/2000 dilution
Predicted band size: 153 kDa
OI-RD Scanning - Anti-MSH6 antibody [EPR3945] (ab92471)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] (ab92471)
Tissue Microarrays stained for "Anti-MSH6 antibody [EPR3945]" using "ab92471"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab92471 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry - Anti-MSH6 antibody [EPR3945] (ab92471)
Immunohistochemical analysis of formalin fixed paraffin embedded human small intestine labelling MSH6 with ab92471 at a concentration of 0.1µg/ml.
The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5. ab92471 anti-MSH6 was incubated at 37°C for 16min.
Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] (ab92471)
Tissue Microarrays stained for "Anti-MSH6 antibody [EPR3945]" using "ab92471"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab92471 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry - Anti-MSH6 antibody [EPR3945] (ab92471)
Immunohistochemical analysis of formalin fixed paraffin embedded human small intestine labelling MSH6 with ab92471 at a concentration of 0.1µg/ml. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.
ab92471 anti-MSH6 antibody [EPR3945] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)