Anti-TMEM175 antibody [EPR29579-563] (ab323710)

Rabbit Recombinant Monoclonal TMEM175 antibody. Suitable for IHC-Fr, IHC-P, WB and reacts with Mouse, Rat samples.

Be the first to review this product! Submit a review

Images

Western blot - Anti-TMEM175 antibody [EPR29579-563] (AB323710), expandable thumbnail

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-Fr	IHC-P	WB	ICC/IF
Human	Not recommended	Not recommended	Not recommended	Not recommended
Mouse	Tested	Tested	Tested	Not recommended
Rat	Tested	Tested	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

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

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

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

Target data

See full target information Tmem175

Function

Proton-activated proton channel that catalyzes proton efflux from endosomes and lysosomes to maintain a steady-state pH (PubMed:35750034). Activated at low pH (under pH 4.6) by luminal side protons: selectively mediates lysosomal proton release from lysosomes, eliciting a proton leak that balances V-ATPase activity to maintain pH homeostasis (By similarity). Regulation of lumenal pH stability is required for autophagosome-lysosome fusion (PubMed:26317472). Also acts as a potassium channel at higher pH, regulating potassium conductance in endosomes and lysosomes (PubMed:26317472, PubMed:33505021). Constitutes the pore-forming subunit of the lysoK(GF) complex, a complex activated by extracellular growth factors (PubMed:33505021). The lysoK(GF) complex is composed of TMEM175 and AKT (AKT1, AKT2 or AKT3), a major target of growth factor receptors: in the complex, TMEM175 channel is opened by conformational changes by AKT, leading to its activation (PubMed:33505021). The lysoK(GF) complex is required to protect neurons against stress-induced damage (PubMed:33505021).

Additional Targets

Tmem175

Alternative names

Endosomal/lysosomal proton channel TMEM175, Potassium channel TMEM175, Transmembrane protein 175, mTMEM175

Rabbit Recombinant Monoclonal TMEM175 antibody. Suitable for IHC-Fr, IHC-P, WB and reacts with Mouse, Rat samples.

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: EPR29579-563
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.

15 product images

Western blot - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: Skeletal muscle
The samples are unboiled.
This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.
The identity of the bands higher than 45 kDa is unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-TMEM175 antibody [EPR29579-563] (ab323710) at 1/1000 dilution
Lane 1: Mouse testis tissue lysate at 20 µg
Lane 2: Mouse midbrain tissue lysate at 20 µg
Lane 3: Mouse lung tissue lysate at 20 µg
Lane 4: Mouse skeletal muscle tissue lysate at 20 µg
Lane 5: Rat testis tissue lysate at 20 µg
Lane 6: Rat midbrain tissue lysate at 20 µg
Lane 7: Rat lung tissue lysate at 20 µg
Lane 8: Rat skeletal muscle tissue 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
Developed using the ECL technique.
Observed band size: 45 kDa, 36 kDa
Exposure time: 125s
Western blot - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The samples are unboiled.
This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.
The identity of the bands higher than 45 kDa is unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-TMEM175 antibody [EPR29579-563] (ab323710) at 1/1000 dilution
Lane 1: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg
Lane 2: MEF (mouse embryo fibroblast) whole cell lysate at 20 µg
Lane 3: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg
Lane 4: C6 (rat glial tumor glial 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
Developed using the ECL technique.
Observed band size: 45 kDa, 36 kDa
Exposure time: 92s
Immunohistochemistry (Frozen sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skeletal muscle (perfused fixed) tissue labeling TMEM175 with ab323710 at 1/50 (9.96 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).
Low expression: confocal image showing no staining on mouse skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab323710 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
Immunohistochemistry (Frozen sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse testis (perfused fixed) tissue labeling TMEM175 with ab323710 at 1/50 (9.96 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).
Confocal image showing positive staining on mouse testis. The nuclear counterstain was DAPI (Blue). The section was incubated with ab323710 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
Immunohistochemistry (Frozen sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skeletal muscle (perfused fixed) tissue labeling TMEM175 with ab323710 at 1/50 (9.96 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).
Low expression: confocal image showing no staining on rat skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab323710 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
Immunohistochemistry (Frozen sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat testis (perfused fixed) tissue labeling TMEM175 with ab323710 at 1/50 (9.96 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).
Confocal image showing positive staining on rat testis. The nuclear counterstain was DAPI (Blue). The section was incubated with ab323710 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling TMEM175 with ab323710 at 1/100 (5.0 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression: no staining on rat skeletal muscle. The section was incubated with Anti-Ki67 [SP6] – Chicken IgY (Chimeric) – BSA and Azide Free ab320710 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling TMEM175 with ab323710 at 1/100 (5.0 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on rat kidney. The section was incubated with Anti-Ki67 [SP6] – Chicken IgY (Chimeric) – BSA and Azide Free ab320710 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling TMEM175 with ab323710 at 1/100 (5.0 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on rat spleen. The section was incubated with Anti-Ki67 [SP6] – Chicken IgY (Chimeric) – BSA and Azide Free ab320710 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling TMEM175 with ab323710 at 1/100 (5.0 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on rat testis. The section was incubated with Anti-Ki67 [SP6] – Chicken IgY (Chimeric) – BSA and Azide Free ab320710 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling TMEM175 with ab323710 at 1/100 (5.0 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression: no staining on mouse skeletal muscle. The section was incubated with Anti-Ki67 [SP6] – Chicken IgY (Chimeric) – BSA and Azide Free ab320710 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling TMEM175 with ab323710 at 1/100 (5.0 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse kidney. The section was incubated with Anti-Ki67 [SP6] – Chicken IgY (Chimeric) – BSA and Azide Free ab320710 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling TMEM175 with ab323710 at 1/100 (5.0 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse spleen. The section was incubated with Anti-Ki67 [SP6] – Chicken IgY (Chimeric) – BSA and Azide Free ab320710 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling TMEM175 with ab323710 at 1/100 (5.0 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse testis. The section was incubated with Anti-Ki67 [SP6] – Chicken IgY (Chimeric) – BSA and Azide Free ab320710 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Western blot - Anti-TMEM175 antibody [EPR29579-563] (ab323710)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The samples are unboiled.
This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-TMEM175 antibody [EPR29579-563] (ab323710) at 1/1000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg
Lane 2: NIH/3T3 transfected with siRNA specifically targeting mouse TMEM175 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
Developed using the ECL technique.
Observed band size: 45 kDa, 36 kDa
Exposure time: 125s