Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)

Rabbit Recombinant Monoclonal ST8SiaIII antibody. Suitable for I-ELISA, ICC/IF, WB, IP, IHC-Fr, IHC-P and reacts with Recombinant fragment - Human, Human, Mouse, Rat samples.

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Images

Immunoprecipitation - Anti-ST8SiaIII antibody [EPR28521-92] (AB317616), 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

	I-ELISA	ICC/IF	WB	IP	IHC-Fr	Flow Cyt	IHC-P
Human	Expected	Tested	Tested	Tested	Expected	Not recommended	Tested
Mouse	Expected	Expected	Tested	Tested	Tested	Not recommended	Tested
Rat	Expected	Expected	Not recommended	Expected	Expected	Not recommended	Tested
Recombinant fragment - Human	Tested	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info 1000 ng/mL	Notes -

Expected
Expected

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

Tested
Tested

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

Expected
Expected

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

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

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

Not recommended
Not recommended

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

Tested
Tested

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

Expected
Expected

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

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes -

Expected
Expected

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

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Not recommended
Not recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	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 Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	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 Recombinant fragment - Human	Dilution info -	Notes -

Target data

See full target information ST8SIA3

Function

Catalyzes the transfer of sialic acid from a CMP-linked sialic acid donor onto a terminal alpha-2,3-, alpha-2,6-, or alpha-2,8-linked sialic acid of an acceptor, such as N-linked oligosaccharides of glycoproteins and glycolipids through alpha-2,8-linkages (PubMed:10766765, PubMed:26192331, PubMed:9826427). Forms oligosialic and polysialic acid on various sialylated N-acetyllactosamine oligosaccharides of glycoproteins, including FETUB N-glycans, a2-HS-glycoprotein (AHSG) and alpha 2,3-sialylated glycosphingolipids, such as alpha 2,3-sialylparagloboside and ganglioside GM3 and to a lesser extent NCAM1 N-glycans (PubMed:10766765, PubMed:9826427). However, it is much more specific to N-linked oligosaccharides of glycoproteins than glycosphingolipids (By similarity). 2,3-sialylparagloboside serves as the best acceptor substrate among the glycolipids (By similarity). alpha-Neu5Ac-(2->8)-alpha-Neu5Ac-(2->3)-beta-D-Gal-(1->4)-6S-D-GlcNAc and monosialyl and disialyl N-acetyllactosamines are the best acceptor substrates among glycoproteins (PubMed:10766765, PubMed:26192331). May plays critical role in the striatum by mediating the formation of disialylated and trisialylated terminal glycotopes on N- and O-glycans of specific striatal proteins, regulating their distribution in lipid rafts, affecting their interaction with other binding partners, and subsequently modulating striatal functions (By similarity).

Alternative names

SIAT8C, ST8SIA3, Ganglioside GD3 synthase ST8SIA3, Sialyltransferase 8C, Sialyltransferase St8Sia III, hST8Sia III, SIAT8-C, ST8SiaIII

Rabbit Recombinant Monoclonal ST8SiaIII antibody. Suitable for I-ELISA, ICC/IF, WB, IP, IHC-Fr, IHC-P and reacts with Recombinant fragment - Human, Human, 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: EPR28521-92
Purification technique: Affinity purification Protein A
Specificity: Unsuitable for rat WB.
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.

Supplementary info

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

Activity summary

ST8SiaIII also known as Sialyltransferase 8B is an enzyme weighing approximately 43 kDa. This enzyme plays a mechanical role in the brain where it contributes to the synthesis of polysialic acid chains. ST8SiaIII adds sialic acid residues to glycoproteins and glycolipids catalyzing the transfer of these residues to terminal ends of N-glycans on proteins notably NCAM1 (Neural Cell Adhesion Molecule 1). The enzyme is expressed in the nervous system specifically detected in the brain tissue where it influences neural activity and development.

Biological function summary

ST8SiaIII modifies neural cell surfaces impacting cell-cell interactions and neural plasticity. It does not operate as a singular entity and is often part of larger glycosylation complexes working in tandem with other sialyltransferases such as ST8SiaII. Through these modifications it regulates the extent of polysialylation which is necessary for processes like cell migration synaptic plasticity and axonal growth. Therefore ST8SiaIII plays a pivotal role in neural development and the central nervous system's adaptability.

Pathways

Polysialylation mediated by ST8SiaIII fits within critical signal transduction pathways like the neural development pathway. This pathway entails multiple proteins where ST8SiaIII interacts particularly with NCAM1 facilitating neuronal changes and synapse formation. It is also part of broader glycosylation pathways contributing to cellular communication and signal transduction. Through these pathways the enzyme influences neural connectivity and cognitive functions forming a network with similar glyco-enzyme participants.

Associated diseases and disorders

ST8SiaIII has associations with neurological conditions such as schizophrenia and Alzheimer’s disease. Changes in polysialic acid expression levels can disrupt neural functions influencing disease progression. These changes affect linked proteins including NCAM1 highlighting the enzyme's role in neural health. Misregulation of ST8SiaIII activity could lead to altered synapse formation and stability therefore contributing to cognitive deficits seen in these disorders. Understanding its function and regulation may provide insights into disease mechanisms and potential therapeutic avenues.

Product promise

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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:
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16 product images

Immunoprecipitation - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
ST8SiaIII was immunoprecipitated from 0.35 mg Mouse striatum tissue lysate with ab317616 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317616 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Mouse striatum tissue lysate

Lane 2: ab317616 IP in Mouse striatum tissue lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab317616 in mouse striatum tissue lysate
All lanes: Immunoprecipitation - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616) at 1/30 dilution
All lanes: Mouse striatum tissue lysate with NFDM/TBST
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 24s
Immunoprecipitation - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
ST8SiaIII was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate with ab317616 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317616 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate

Lane 2: ab317616 IP in SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab317616 in SH-SY5Y whole cell lysate
All lanes: Immunoprecipitation - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616) at 1/30 dilution
All lanes: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate with NFDM/TBST
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 120s
Immunocytochemistry/ Immunofluorescence - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling ST8SiaIII with ab317616 at 1/50 (10.58 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 antibody at 1/1000 2 ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in SH-SY5Y cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).

Low expression: U-937.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only 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.
Western blot - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
Low expression: U-937.
ST8SiaIII is a glycoprotein of approximately 45-60 kDa and detected as a 37-40 kDa band after treated with Protein Deglycosylation MIX II.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 11980897, PMID: 10766765).
The identity of the higher MW band at approximately 75 kDa is unknown.
Samples are non-boiled as boiling may cause protein aggregation.
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-ST8SiaIII antibody [EPR28521-92] (ab317616) at 1/1000 dilution
Lane 1: Un treated SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST
Lane 2: SH-SY5Y cell lysate treated with Peptide:N-glycosidase F (PNGase F) at 20 µg with NFDM/TBST
Lane 3: U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg with NFDM/TBST
Lane 4: SK-N-BE(2) (human neuroblastoma neuroblast) whole cell lysate at 20 µg with NFDM/TBST
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: 45-60 kDa, 37-40 kDa, 36 kDa
Exposure time: 180s
Indirect ELISA - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
Indirect ELISA analysis of ab317616 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1:2500 dilution.
Antigen: Human ST8SiaIII.
Antigen concentration: 1000 ng/ml
Immunohistochemistry (Frozen sections) - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh frozen) tissue labeling ST8SiaIII with ab317616 at 1/100 (5.29 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).
Negative control: confocal image showing no staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317616 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) preadsorbedat 1/1000 2 ug/mL dilution.
Immunohistochemistry (Frozen sections) - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh frozen) tissue labeling ST8SiaIII with ab317616 at 1/100 (5.29 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).
Panel A: merged staining of anti-ST8SiaIII (ab317616, green), anti-NeuN (Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, grey) and anti-GFAP (Alexa Fluor® 594 Anti-GFAP antibody [EPR1034Y] ab201732, magenta) on mouse cerebrum.

Panel B: anti-ST8SIA3 stained on mouse cerebrum.

Panel C: anti-NeuN stained in neurons of mouse cerebrum.

Panel D: anti-GFAP stained in astrocytes of mouse cerebrum.

The section was incubated in two rounds of staining: in the order of ab317616 and Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, Alexa Fluor® 594 Anti-GFAP antibody [EPR1034Y] ab201732 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). 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) preadsorbedat 1/1000 2 ug/mL dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling ST8SiaIII with ab317616 at 1/1000 (0.529 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: No staining on rat liver.

The section was incubated with ab317616 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-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling ST8SiaIII with ab317616 at 1/1000 (0.529 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: No staining on mouse liver.

The section was incubated with ab317616 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-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling ST8SiaIII with ab317616 at 1/100 (5.29 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: No staining on human liver.

The section was incubated with ab317616 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-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling ST8SiaIII with ab317616 at 1/1000 (0.529 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on rat cerebrum.

The section was incubated with ab317616 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-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling ST8SiaIII with ab317616 at 1/1000 (0.529 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse cerebrum (PMID: 7782326).

The section was incubated with ab317616 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-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling ST8SiaIII with ab317616 at 1/100 (5.29 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human cerebellum.

The section was incubated with ab317616 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-ST8SiaIII antibody [EPR28521-92] (ab317616)
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling ST8SiaIII with ab317616 at 1/100 (5.29 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human cerebrum (PMID: 9826427).

The section was incubated with ab317616 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-ST8SiaIII antibody [EPR28521-92] (ab317616)
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-ST8SiaIII antibody [EPR28521-92] (ab317616) at 1/1000 dilution
All lanes: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg with NFDM/TBST
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: 45 kDa, 36 kDa
Exposure time: 180s
Western blot - Anti-ST8SiaIII antibody [EPR28521-92] (ab317616)
Negative control: liver, placenta, lung (PMID: 9826427, PMID: 7782326).
Samples are non-boiled as boiling may cause protein aggregation.
The identity of the higher MW band at approximately 75 kDa (in lanes 1-6 human tissues) is unknown.
The identity of the lower MW band at approximately 37 kDa and higher MW band at approximately 150 kDa (in lane 11) is unknown.
Lanes 7-12 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 and lanes 1-6 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.
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-ST8SiaIII antibody [EPR28521-92] (ab317616) at 1/1000 dilution
Lane 1: Human hippocampus tissue lysate at 20 µg with NFDM/TBST
Lane 2: Human hypothalamus tissue lysate at 20 µg with NFDM/TBST
Lane 3: Human brain tissue lysate at 20 µg with NFDM/TBST
Lane 4: Human striatum tissue lysate at 20 µg with NFDM/TBST
Lane 5: Human liver tissue lysate at 20 µg with NFDM/TBST
Lane 6: Human placenta tissue lysate at 20 µg with NFDM/TBST
Lane 7: Mouse brain tissue lysate at 20 µg with NFDM/TBST
Lane 8: Mouse hippocampus tissue lysate at 20 µg with NFDM/TBST
Lane 9: Mouse striatum tissue lysate at 20 µg with NFDM/TBST
Lane 10: Mouse cerebellum tissue lysate at 20 µg with NFDM/TBST
Lane 11: Mouse liver tissue lysate at 20 µg with NFDM/TBST
Lane 12: Mouse lung tissue lysate at 20 µg with NFDM/TBST
Secondary
Lanes 1 - 6: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Lanes 7 - 12: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 45 kDa, 36 kDa
Exposure time: 180s