Anti-CMTM6 antibody [EPR23015-45] (ab264067)

Rabbit Recombinant Monoclonal CMTM6 antibody. Suitable for WB, Flow Cyt (Intra), IHC-P, IP and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

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Images

Western blot - Anti-CMTM6 antibody [EPR23015-45] (AB264067), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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

	WB	ICC/IF	Flow Cyt (Intra)	IHC-P	IP
Human	Tested	Not recommended	Tested	Tested	Expected
Mouse	Tested	Not recommended	Expected	Tested	Tested
Rat	Tested	Not recommended	Expected	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -

Not recommended
Not recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes -
Species Rat	Dilution info 1/30	Notes -

Expected
Expected

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

Associated Products

Select an associated product type

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Target data

See full target information CMTM6

Function

Master regulator of recycling and plasma membrane expression of PD-L1/CD274, an immune inhibitory ligand critical for immune tolerance to self and antitumor immunity. Associates with both constitutive and IFNG-induced PD-L1/CD274 at recycling endosomes, where it protects PD-L1/CD274 from being targeted for lysosomal degradation, likely by preventing its STUB1-mediated ubiquitination. May stabilize PD-L1/CD274 expression on antigen presenting cells and potentiates inhibitory signaling by PDCD1/CD279, its receptor on T-cells, ultimately triggering T-cell anergy.

Alternative names

CKLFSF6, CMTM6, CKLF-like MARVEL transmembrane domain-containing protein 6, Chemokine-like factor superfamily member 6

Recommended products

Rabbit Recombinant Monoclonal CMTM6 antibody. Suitable for WB, Flow Cyt (Intra), IHC-P, IP and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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: EPR23015-45
Purification technique: Affinity purification Protein A
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: Avoid freeze / thaw cycle

Notes

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.

11 product images

Western blot - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lanes 1-2: 3 minutes
Lane 3: 48 seconds.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 28813417).
All lanes: Western blot - Anti-CMTM6 antibody [EPR23015-45] (ab264067) at 1/1000 dilution
Lane 1: MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2: Human tonsil tissue lysate at 20 µg
Lane 3: BxPC-3 (human pancreas adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 20 kDa
Observed band size: 20 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Immunohistochemical analysis of paraffin-embedded Human pancreatic cancer tissue labeling CMTM6 with ab264067 at 1/2000 dilution (0.24 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Membranous staining on human pancreatic cancer (PMID/ 28813417) The section was incubated with ab264067 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control/ Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Immunohistochemical analysis of paraffin-embedded Human thyroid cancer tissue labeling CMTM6 with ab264067 at 1/2000 dilution (0.24 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Membranous staining on human thyroid cancer (PMID: 28813410) The section was incubated with ab264067 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND™ RX instrument Counterstained with Hematoxylin.
Secondary antibody only control/ Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CMTM6 with ab264067 at 1/2000 dilution (0.24 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human tonsil. The section was incubated with ab264067 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control/ Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Western blot - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Exposure time:
Lane 1-2: 15 seconds
Lane 3-5: 10 seconds

Blocking/Diluting buffer and concentration 5% NFDM/TBST

Loading control: Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) (1/200000) (36KDa).

Low expression: skeletal muscle.
This blot was developed using a high sensitivity ECL substrate.
Samples are non-boiled as boiling may cause protein aggregates.
Lanes 1 - 5: Western blot - Anti-CMTM6 antibody [EPR23015-45] (ab264067) at 1/1000 dilution
Lanes 1 - 5: Western blot - Anti-CMTM6 antibody [EPR23015-45] - BSA and Azide free (Anti-CMTM6 antibody [EPR23015-45] - BSA and Azide free ab264076)
Lane 1: Mouse spleen tissue lysate at 20 µg
Lane 2: Mouse skeletal muscle tissue lysate at 20 µg
Lane 3: Rat spleen tissue lysate at 20 µg
Lane 4: Rat skeletal muscle tissue lysate at 20 µg
Lane 5: Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling CMTM6 with ab264067 at 1/2000 dilution, incubated 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). Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Positive staining on mouse testis.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Flow Cytometry (Intracellular) - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized MDA-MB-231 (human breast adenocarcinoma epithelial cell) cells labelling CMTM6 with ab264067 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)isotype control (black)and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Blocking and diluting buffer and concentration: 5% NFDM /TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.

We recommend not to boil the samples after lysis to get desired WB bands.
All lanes: Western blot - Anti-CMTM6 antibody [EPR23015-45] (ab264067) at 1/10000 dilution
Lane 1: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate boiled at 20 µg
Lane 2: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate unboiled at 20 µg
Lane 3: BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate boiled at 20 µg
Lane 4: BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate unboiled at 20 µg
Lane 5: Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate boiled at 20 µg
Lane 6: Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate unboiled at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 20 kDa
Observed band size: 20 kDa
Exposure time: 7s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling CMTM6 with ab264067 at 1/2000 dilution, incubated 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). Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Low expression: nearly no staining on mouse skeletal muscle.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Immunoprecipitation - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
ab264067 at 1/30 dilution immunoprecipitating CMTM6 in mouse spleen tissue lysate. Western blot was performed from the immunoprecipitate using ab264067 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.

Lane 1: Mouse spleen tissue lysate 10 µg

Lane 2: ab264067 IP in mouse spleen tissue lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab264067 in mouse spleen tissue lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 8 seconds.
Lane 2: Immunoprecipitation - Anti-CMTM6 antibody [EPR23015-45] (ab264067) at 1/30 dilution
Lane 2: Immunoprecipitation - Anti-CMTM6 antibody [EPR23015-45] - BSA and Azide free (Anti-CMTM6 antibody [EPR23015-45] - BSA and Azide free ab264076)
All lanes: Mouse spleen tissue lysate at 10 µg
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 8s
Immunoprecipitation - Anti-CMTM6 antibody [EPR23015-45] (ab264067)
ab264067 at 1/30 dilution immunoprecipitating CMTM6 in rat spleen tissue lysate. Western blot was performed from the immunoprecipitate using ab264067 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.

Lane 1: Rat spleen tissue lysate 10 µg

Lane 2: ab264067 IP in rat spleen tissue lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab264067 in rat spleen tissue lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 6 seconds.
Lane 2: Immunoprecipitation - Anti-CMTM6 antibody [EPR23015-45] (ab264067) at 1/30 dilution
Lane 2: Immunoprecipitation - Anti-CMTM6 antibody [EPR23015-45] - BSA and Azide free (Anti-CMTM6 antibody [EPR23015-45] - BSA and Azide free ab264076)
All lanes: Mouse spleen tissue lysate at 10 µg
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 6s