Name: Anti-TIA1 antibody [EPR9304]
SKU: ab140595
Rating: 2 (1 reviews)

Anti-TIA1 antibody [EPR9304] (ab140595) is a rabbit monoclonal antibody that is used to detect TIA1 in Western Blot, IP, IHC-P, ICC/IF. Suitable for Human, Mouse samples.

- Specificity confirmed with TIA1 knockout cell line validation

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Images

Immunocytochemistry/ Immunofluorescence - Anti-TIA1 antibody [EPR9304] (AB140595), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: 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.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	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 Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes For unpurified use at 1/100 - 1/250. 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 Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information TIA1

Function

RNA-binding protein involved in the regulation of alternative pre-RNA splicing and mRNA translation by binding to uridine-rich (U-rich) RNA sequences (PubMed:11106748, PubMed:12486009, PubMed:17488725, PubMed:8576255). Binds to U-rich sequences immediately downstream from a 5' splice sites in a uridine-rich small nuclear ribonucleoprotein (U snRNP)-dependent fashion, thereby modulating alternative pre-RNA splicing (PubMed:11106748, PubMed:8576255). Preferably binds to the U-rich IAS1 sequence in a U1 snRNP-dependent manner; this binding is optimal if a 5' splice site is adjacent to IAS1 (By similarity). Activates the use of heterologous 5' splice sites; the activation depends on the intron sequence downstream from the 5' splice site, with a preference for a downstream U-rich sequence (PubMed:11106748). By interacting with SNRPC/U1-C, promotes recruitment and binding of spliceosomal U1 snRNP to 5' splice sites followed by U-rich sequences, thereby facilitating atypical 5' splice site recognition by U1 snRNP (PubMed:11106748, PubMed:12486009, PubMed:17488725). Activates splicing of alternative exons with weak 5' splice sites followed by a U-rich stretch on its own pre-mRNA and on TIAR mRNA (By similarity). Acts as a modulator of alternative splicing for the apoptotic FAS receptor, thereby promoting apoptosis (PubMed:11106748, PubMed:17488725, PubMed:1934064). Binds to the 5' splice site region of FAS intron 5 to promote accumulation of transcripts that include exon 6 at the expense of transcripts in which exon 6 is skipped, thereby leading to the transcription of a membrane-bound apoptotic FAS receptor, which promotes apoptosis (PubMed:11106748, PubMed:17488725, PubMed:1934064). Binds to a conserved AU-rich cis element in COL2A1 intron 2 and modulates alternative splicing of COL2A1 exon 2 (PubMed:17580305). Also binds to the equivalent AT-rich element in COL2A1 genomic DNA, and may thereby be involved in the regulation of transcription (PubMed:17580305). Binds specifically to a polypyrimidine-rich controlling element (PCE) located between the weak 5' splice site and the intronic splicing silencer of CFTR mRNA to promote exon 9 inclusion, thereby antagonizing PTB1 and its role in exon skipping of CFTR exon 9 (PubMed:14966131). Involved in the repression of mRNA translation by binding to AU-rich elements (AREs) located in mRNA 3' untranslated regions (3' UTRs), including target ARE-bearing mRNAs encoding TNF and PTGS2 (By similarity). Also participates in the cellular response to environmental stress, by acting downstream of the stress-induced phosphorylation of EIF2S1/EIF2A to promote the recruitment of untranslated mRNAs to cytoplasmic stress granules (SGs), leading to stress-induced translational arrest (PubMed:10613902). Formation and recruitment to SGs is regulated by Zn(2+) (By similarity). Possesses nucleolytic activity against cytotoxic lymphocyte target cells (PubMed:1934064). Isoform Short. Displays enhanced splicing regulatory activity compared with TIA isoform Long.

Alternative names

Cytotoxic granule associated RNA binding protein TIA1, Nucleolysin TIA-1 isoform p40, RNA-binding protein TIA-1, T-cell-restricted intracellular antigen-1, p40-TIA-1, TIA-1, TIA1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: 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: EPR9304
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

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

What are the advantages of a recombinant monoclonal antibody?
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.

Species reactivity
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.

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

Immunocytochemistry/ Immunofluorescence - Anti-TIA1 antibody [EPR9304] (ab140595)
ab140595 staining TIA1 in wild-type Hap1 cells (top panel) and TIA1 knockout Hap1 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (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 with ab140595 at 1/250 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution 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) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR9304] (ab140595)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling TIA1 with purified ab140595 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-TIA1 antibody [EPR9304] (ab140595)
Immunocytochemistry/Immunofluorescence analysis of HuT-78 cells labelling TIA1 with purified ab140595 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).
Western blot - Anti-TIA1 antibody [EPR9304] (ab140595)
Lane 1: Wild-type HAP1 cell lysate (40 μg)
Lane 2: TIA1 knockout HAP1 cell lysate (40 μg)
Lane 3: Jurkat cell lysate (40 μg)
Lane 4: K562 cell lysate (40 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab140595 observed at 43 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab140595 was shown to specifically react with TIA1 when TIA1 knockout samples were used. Wild-type and TIA1 knockout samples were subjected to SDS-PAGE. ab140595 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10000 dilution respectively and incubated overnight at 4C. 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 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-TIA1 antibody [EPR9304] (ab140595)
Predicted band size: 43 kDa
Western blot - Anti-TIA1 antibody [EPR9304] (ab140595)
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-TIA1 antibody [EPR9304] (ab140595) at 1/5000 dilution
All lanes: NIH/3T3 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 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 42-43 kDa
Immunoprecipitation - Anti-TIA1 antibody [EPR9304] (ab140595)
ab140595 (purified) at 1/40 immunoprecipitating TIA1 in HuT-78 whole cell lysate.
Lane 1 (input): HuT-78 whole cell lysate (10μg)
Lane 2 (+): ab140595 + HuT-78 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab140595 in HuT-78 whole cell lysate.
For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Immunoprecipitation - Anti-TIA1 antibody [EPR9304] (ab140595)
Predicted band size: 43 kDa
Observed band size: 42 kDa, 43 kDa
Western blot - Anti-TIA1 antibody [EPR9304] (ab140595)
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-TIA1 antibody [EPR9304] (ab140595) at 1/5000 dilution
Lane 1: Jurkat whole cell lysate at 20 µg
Lane 2: K562 whole cell lysate at 20 µg
Lane 3: HUT-78 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 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 42-43 kDa
Western blot - Anti-TIA1 antibody [EPR9304] (ab140595)
All lanes: Western blot - Anti-TIA1 antibody [EPR9304] (ab140595)
Lane 1: HuT-78 cell lysate at 10 µg
Lane 2: Jurkat cell lysate at 10 µg
Lane 3: Molt-4 cell lysate at 10 µg
Lane 4: K562 cell lysate at 10 µg
Secondary
All lanes: HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 43 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR9304] (ab140595)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling TIA1 with unpurified ab140595 at a dilution of 1/100.
Immunoprecipitation - Anti-TIA1 antibody [EPR9304] (ab140595)
Immunoprecipitation of TIA1 in HAP1 cells. Lysates were prepared and immunoprecipitation was performed using 1.0 µg of Anti-TIA1 antibody [EPR22999-80] ab263945 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Immunoprecipitation - Anti-TIA1 antibody [EPR9304] (ab140595) at 1 µg
All lanes: HAP1 cells
Western blot - Anti-TIA1 antibody [EPR9304] (ab140595)
ab140595 was shown to react with TIA1 in wild-type HAP1 cells in Western blot with loss of signal observed in a TIA1 knockout cell line. Wild-type HAP1 and TIA1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab140595 overnight at 4 °C at a 1/5000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-TIA1 antibody [EPR9304] (ab140595) at 1/5000 dilution
Lane 1: Wild-type HAP1 lysate at 50 µg
Lane 2: TIA1 knock-out HAP1 lysate at 50 µg