Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

Rabbit Recombinant Monoclonal MSH6 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB and reacts with Rat, Human, Mouse samples. Cited in 9 publications.

Be the first to review this product! Submit a review

Related conjugates and formulations

ab311007
Conjugated
Alexa Fluor® 488 Anti-MSH6 antibody [EPR3945]
ab92471
Unconjugated
Anti-MSH6 antibody [EPR3945]
ab321758
Conjugated
Alexa Fluor® 750 Anti-MSH6 antibody [EPR3945]
ab198334
Conjugated
Alexa Fluor® 647 Anti-MSH6 antibody [EPR3945]
ab311771
Conjugated
Alexa Fluor® 594 Anti-MSH6 antibody [EPR3945]
ab313252
Conjugated
Alexa Fluor® 555 Anti-MSH6 antibody [EPR3945]

Images

Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (AB214454), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
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	ICC/IF	WB
Human	Tested	Tested	Tested
Mouse	Expected	Expected	Expected
Rat	Tested	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat, Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

Select an associated product type

6 products for Alternative Product

3 products for Alternative Version

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

Recommended products

Rabbit Recombinant Monoclonal MSH6 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB and reacts with Rat, Human, Mouse samples. Cited in 9 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
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 conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab214454 is the carrier-free version of Anti-MSH6 antibody [EPR3945] ab92471.

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

MSH6 also known as MutS Homolog 6 is a DNA repair protein that plays a role in the mismatch repair (MMR) system. It has a molecular mass of approximately 136 kDa. MSH6 forms a heterodimer with MSH2 called MutSα and this complex identifies base-pair mismatches and insertion-deletion loops during DNA replication. It is expressed in various tissues throughout the body and high levels are often found in proliferative tissues where active DNA replication occurs.

Biological function summary

MSH6 functions as part of the MMR complex which is essential for maintaining genomic stability. The MutSα complex where MSH6 pairs with MSH2 operates along with other proteins in the MMR pathway to correct DNA replication errors. MSH6 is also known to interact with PCNA a DNA polymerase processivity factor which facilitates its role in the repair process.

Pathways

MSH6 participates prominently in the DNA mismatch repair pathway. This pathway is critical for correcting DNA errors and preventing mutations during replication. In association with MLH1-PMS2 (MutLα complex) MSH6 ensures that DNA integrity is preserved. Additionally MSH6 is involved in the base excision repair (BER) pathway where it collaborates with other repair proteins to fix small base lesions.

Associated diseases and disorders

MSH6 has a significant connection with Lynch syndrome also known as hereditary nonpolyposis colorectal cancer (HNPCC). This condition is characterized by germline mutations in MMR genes including MSH6 leading to increased cancer risk particularly in the colon. Moreover alterations in MSH6 can contribute to microsatellite instability a feature seen in certain types of endometrial cancer. Mutations in MSH2 often accompany MSH6 mutations in these disorders further impacting the MMR pathway's efficiency.

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.

14 product images

Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Clone EPR3945 (ab214454) has been successfully conjugated by Abcam. This image was generated using Anti-MSH6 antibody [EPR3945] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-MSH6 antibody [EPR3945] ab198334 for protocol details.
Alexa Fluor® 647 Anti-MSH6 antibody [EPR3945] ab198334 staining MSH6 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), 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 overnight at +4°C with Alexa Fluor® 647 Anti-MSH6 antibody [EPR3945] ab198334 at 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Immunohistochemical staining of paraffin embedded human colon with unpurified Anti-MSH6 antibody [EPR3945] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH6 antibody [EPR3945] ab92471).
Western blot - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
This data was developed using the same antibody clone in a different buffer formulation (Anti-MSH6 antibody [EPR3945] ab92471).
Lanes 1- 2: Merged signal (red and green). Green - Anti-MSH6 antibody [EPR3945] ab92471 observed at 160 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
Anti-MSH6 antibody [EPR3945] 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. Anti-MSH6 antibody [EPR3945] 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] (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] - BSA and Azide free (ab214454)
Anti-MSH6 antibody [EPR3945] 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 Anti-MSH6 antibody [EPR3945] 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH6 antibody [EPR3945] ab92471).
Western blot - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
This data was developed using Anti-MSH6 antibody [EPR3945] ab92471, the same antibody clone in a different buffer formulation. Different batches of Anti-MSH6 antibody [EPR3945] 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] (Anti-MSH6 antibody [EPR3945] ab92471)
Predicted band size: 153 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Immunohistochemical staining of paraffin embedded rat liver with purified Anti-MSH6 antibody [EPR3945] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH6 antibody [EPR3945] ab92471).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Immunohistochemical staining of paraffin embedded rat liver with unpurified Anti-MSH6 antibody [EPR3945] ab92471 at a dilution of 1/150. 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH6 antibody [EPR3945] ab92471).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Immunohistochemical staining of paraffin embedded human colon with unpurified Anti-MSH6 antibody [EPR3945] ab92471 at a dilution of 1/150. 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH6 antibody [EPR3945] ab92471).
Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Anti-MSH6 antibody [EPR3945] 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 Anti-MSH6 antibody [EPR3945] 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH6 antibody [EPR3945] ab92471).
Image courtesy of a customer review submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Unpurified Anti-MSH6 antibody [EPR3945] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH6 antibody [EPR3945] ab92471).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Unpurified Anti-MSH6 antibody [EPR3945] ab92471, at a 1/100 dilution, detecting MSH6 in paraffin embedded Human colonic adenocarcinoma tissue by immunohistochemistry. Detection used HRP conjugated anti rabbit antibody.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH6 antibody [EPR3945] ab92471).
OI-RD Scanning - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
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] - BSA and Azide free (ab214454)
Tissue Microarrays stained for "Anti-MSH6 antibody [EPR3945]" using "Anti-MSH6 antibody [EPR3945] 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 Anti-MSH6 antibody [EPR3945] 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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
Tissue Microarrays stained for "Anti-MSH6 antibody [EPR3945]" using "Anti-MSH6 antibody [EPR3945] 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 Anti-MSH6 antibody [EPR3945] 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.