Name: Anti-TFEB antibody [EPR22941-6]
SKU: ab267351
Rating: 5 (1 reviews)

Rabbit Recombinant Monoclonal TFEB antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 4 publications.

Images

Immunoprecipitation - Anti-TFEB antibody [EPR22941-6] (AB267351), 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

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

Tested
Tested

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

Tested
Tested

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

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Tested
Tested

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

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Tested
Tested

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

Associated Products

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Target data

See full target information TFEB

Function

Transcription factor that acts as a master regulator of lysosomal biogenesis, autophagy, lysosomal exocytosis, lipid catabolism, energy metabolism and immune response (PubMed:21617040, PubMed:22343943, PubMed:22576015, PubMed:22692423, PubMed:25720963, PubMed:30120233, PubMed:31672913, PubMed:32612235, PubMed:32753672, PubMed:35662396, PubMed:36697823, PubMed:36749723, PubMed:37079666). Specifically recognizes and binds E-box sequences (5'-CANNTG-3'); efficient DNA-binding requires dimerization with itself or with another MiT/TFE family member such as TFE3 or MITF (PubMed:1748288, PubMed:19556463, PubMed:29146937). Involved in the cellular response to amino acid availability by acting downstream of MTOR: in the presence of nutrients, TFEB phosphorylation by MTOR promotes its cytosolic retention and subsequent inactivation (PubMed:21617040, PubMed:22343943, PubMed:22576015, PubMed:22692423, PubMed:25720963, PubMed:32612235, PubMed:32753672, PubMed:35662396, PubMed:36697823). Upon starvation or lysosomal stress, inhibition of MTOR induces TFEB dephosphorylation, resulting in nuclear localization and transcription factor activity (PubMed:22343943, PubMed:22576015, PubMed:22692423, PubMed:25720963, PubMed:32612235, PubMed:32753672, PubMed:35662396, PubMed:36697823). Specifically recognizes and binds the CLEAR-box sequence (5'-GTCACGTGAC-3') present in the regulatory region of many lysosomal genes, leading to activate their expression, thereby playing a central role in expression of lysosomal genes (PubMed:19556463, PubMed:22692423). Regulates lysosomal positioning in response to nutrient deprivation by promoting the expression of PIP4P1 (PubMed:29146937). Acts as a positive regulator of autophagy by promoting expression of genes involved in autophagy (PubMed:21617040, PubMed:22576015, PubMed:23434374, PubMed:27278822). In association with TFE3, activates the expression of CD40L in T-cells, thereby playing a role in T-cell-dependent antibody responses in activated CD4(+) T-cells and thymus-dependent humoral immunity (By similarity). Specifically recognizes the gamma-E3 box, a subset of E-boxes, present in the heavy-chain immunoglobulin enhancer (PubMed:2115126). Plays a role in the signal transduction processes required for normal vascularization of the placenta (By similarity). Involved in the immune response to infection by the bacteria S.aureus, S.typhimurium or S.enterica: infection promotes itaconate production, leading to alkylation, resulting in nuclear localization and transcription factor activity (PubMed:35662396). Itaconate-mediated alkylation activates TFEB-dependent lysosomal biogenesis, facilitating the bacteria clearance during the antibacterial innate immune response (PubMed:35662396). In association with ACSS2, promotes the expression of genes involved in lysosome biogenesis and both autophagy upon glucose deprivation (PubMed:28552616).

Alternative names

BHLHE35, TFEB, Transcription factor EB, Class E basic helix-loop-helix protein 35, bHLHe35

Recommended products

Rabbit Recombinant Monoclonal TFEB antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 4 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: EPR22941-6
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.

12 product images

Immunoprecipitation - Anti-TFEB antibody [EPR22941-6] (ab267351)
TFEB was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab267351 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267351 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate 10ug
Lane 2: ab267351 IP in Raji whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab267351 in Raji whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds.
The weak band around 50kDa is one isoform of TFEB(PMID:28993453).
All lanes: Immunoprecipitation - Anti-TFEB antibody [EPR22941-6] (ab267351)
Predicted band size: 53 kDa
Observed band size: 66 kDa
Immunoprecipitation - Anti-TFEB antibody [EPR22941-6] (ab267351)
TFEB was immunoprecipitated from 0.35 mg IM-9 (human multiple myeloma B Lymphoblast) whole cell lysate with ab267351 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267351 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: IM-9 (human multiple myeloma B Lymphoblast) whole cell lysate 10ug
Lane 2: ab267351 IP in IM-9 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab267351 in IM-9 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 2 min.
The weak band around 50kDa is one isoform of TFEB(PMID:28993453).
All lanes: Immunoprecipitation - Anti-TFEB antibody [EPR22941-6] (ab267351)
Predicted band size: 53 kDa
Observed band size: 66 kDa
Western blot - Anti-TFEB antibody [EPR22941-6] (ab267351)
False colour image of Western blot: Anti-TFEB antibody [EPR22941-6] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab267351 was shown to bind specifically to TFEB. A band was observed at 60 kDa in wild-type HepG2 cell lysates with no signal observed at this size in TFEB knockout cell line Human TFEB knockout Hep G2 cell line ab277871 (knockout cell lysate ab284220). To generate this image, wild-type and TFEB knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-TFEB antibody [EPR22941-6] (ab267351) at 1/1000 dilution
Lane 1: Wild-type HepG2 cell lysate at 20 µg
Lane 2: TFEB knockout HepG2 cell lysate at 20 µg
Lane 2: Western blot - Human TFEB knockout Hep G2 cell line (Human TFEB knockout Hep G2 cell line ab277871)
Lane 3: Raji cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 53 kDa
Observed band size: 60 kDa
Western blot - Anti-TFEB antibody [EPR22941-6] (ab267351)
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
The molecular weight observed is consistent with what has been described in the literature (PMID:30519051).
All lanes: Western blot - Anti-TFEB antibody [EPR22941-6] (ab267351) at 1/1000 dilution
Lane 1: Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 10 µg
Lane 2: IM-9 (human multiple myeloma B Lymphoblast whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 53 kDa
Observed band size: 66 kDa
Immunocytochemistry/ Immunofluorescence - Anti-TFEB antibody [EPR22941-6] (ab267351)
Immunofluorescent analysis of 100% Methanol-fixed IM-9 (human multiple myeloma B Lymphoblast) cells labelling TFEB with ab267351 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in IM-9 cells. 100% methanol fixation is recommended. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-TFEB antibody [EPR22941-6] (ab267351)
Immunofluorescent analysis of 100% Methanol-fixed Raji (human Burkitt's lymphoma B lymphocyte) cells labelling TFEB with ab267351 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Raji cells. 100% methanol fixation is recommended. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TFEB antibody [EPR22941-6] (ab267351)
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling TFEB with ab267351 at 1/500 dilution (1.042ug/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 in human breast cancer (PMID:28017540). The section was incubated with Anti-C Reactive Protein antibody [EPR22862-10] ab256492 for 15 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-TFEB antibody [EPR22941-6] (ab267351)
Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling TFEB with ab267351 at 1/500 dilution (1.042ug/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 in human lung cancer (PMID: 26264650) The section was incubated with Anti-C Reactive Protein antibody [EPR22862-10] ab256492 for 15 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-TFEB antibody [EPR22941-6] (ab267351)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling TFEB with ab267351 at 1/500 dilution (1.042ug/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 human colon (PMID:30519051). The section was incubated with Anti-C Reactive Protein antibody [EPR22862-10] ab256492 for 15 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-TFEB antibody [EPR22941-6] (ab267351)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: MCF7.
The lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, anti-TFEB antibody (ab267351) staining at 1/1000 dilution.Anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Exposure time: 81 seconds
All lanes: Western blot - Anti-TFEB (phospho S211) antibody [EPR27960-64] (Anti-TFEB (phospho S211) antibody [EPR27960-64] ab310331) at 1/1000 dilution
Lane 1: Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate (untreated membrane) at 20 µg
Lane 2: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg
Lane 3: Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate (phosphatase treated membrane) at 20 µg
Lane 4: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate (phosphatase treated membrane) 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
Observed band size: 70 kDa
Exposure time: 81s
Western blot - Anti-TFEB antibody [EPR22941-6] (ab267351)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression of TFEB (phospho S211) is downregulated in response to Torin 1 treatment (PMID: 28055300).

In Western blot, anti--TFEB antibody (ab267351) staining at 1/1000 dilution.Anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Exposure time: 81 seconds
All lanes: Western blot - Anti-TFEB (phospho S211) antibody [EPR27960-64] (Anti-TFEB (phospho S211) antibody [EPR27960-64] ab310331) at 1/1000 dilution
Lane 1: Untreated Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg
Lane 2: Raji treated with 250 nM Torin 1 for 5 hours 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
Observed band size: 70 kDa
Exposure time: 81s
Flow Cytometry (Intracellular) - Anti-TFEB antibody [EPR22941-6] (ab267351)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized IM-9 (human multiple myeloma B Lymphoblast) cells labelling TFEB with ab267351 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control 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.