Name: Anti-SPR antibody [EPR9290]
Brand: RabMAb
SKU: ab157194
Rating: 4 (1 reviews)

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Western blot - Anti-SPR antibody [EPR9290] (ab157194)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR9290] to SPR
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
Knockout validated
Reacts with: Human

Carrier Free

Product name

Anti-SPR antibody [EPR9290]
See all SPR primary antibodies
Description

Rabbit monoclonal [EPR9290] to SPR
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: A549, MCF7, HepG2 and HeLa (ab150035) cell lysates. IHC-P: Human thyroid gland carcinoma tissue. ICC/IF: HeLa cells. Flow Cyt (intra): MCF7 cells.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at -20ºC.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR9290
Isotype

IgG
Research areas

Alternative Versions
- Anti-SPR antibody [EPR9290] - BSA and Azide free (ab249302)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human SPR knockout HeLa cell line (ab264681)
KO cell lysates
- Human SPR knockout HeLa cell lysate (ab258700)
Positive Controls
Recombinant Protein
- Recombinant Mouse SPR protein (His tag) (ab226880)
Related Products
- Recombinant Human SPR protein (ab86445)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab157194 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB	(1)	1/1000 - 1/10000. Predicted molecular weight: 28 kDa.
IHC-P		1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF		1/100 - 1/250.
IP		1/10 - 1/100.

Notes
Flow Cyt (Intra) 1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000 - 1/10000. Predicted molecular weight: 28 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
IP 1/10 - 1/100.

Function

Catalyzes the final one or two reductions in tetra-hydrobiopterin biosynthesis to form 5,6,7,8-tetrahydrobiopterin.
Involvement in disease

Defects in SPR are the cause of dystonia DOPA-responsive due to sepiapterin reductase deficiency (DRDSPRD) [MIM:612716]. In the majority of cases, patients manifest progressive psychomotor retardation, dystonia and spasticity. Cognitive anomalies are also often present. The disease is due to severe dopamine and serotonin deficiencies in the central nervous system caused by a defect in BH4 synthesis. Dystonia is defined by the presence of sustained involuntary muscle contractions, often leading to abnormal postures.
Sequence similarities

Belongs to the sepiapterin reductase family.
Post-translational
modifications

In vitro phosphorylation of Ser-213 by CaMK2 does not change kinetic parameters.
Cellular localization

Cytoplasm.
Target information above from: UniProt accession P35270 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 6697 Human
- Omim: 182125 Human
- SwissProt: P35270 Human
- Unigene: 301540 Human
Alternative names
- OTTHUMP00000160199 antibody
- SDR38C1 antibody
- Sepiapterin reductase (7,8 dihydrobiopterin:NADP+ oxidoreductase) antibody
- Sepiapterin reductase antibody
- Short chain dehydrogenase/reductase family 38C, member 1 antibody
- SPR antibody
- SPRE_HUMAN antibody
see all

Western blot - Anti-SPR antibody [EPR9290] (ab157194)

All lanes : Anti-SPR antibody [EPR9290] (ab157194) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SPR knockout HeLa cell lysate
Lane 3 : MCF7 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 28 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab157194 observed at 26 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab157194 Anti-SPR antibody [EPR9290] was shown to specifically react with SPR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264681 (knockout cell lysate ab258700) was used. Wild-type and SPR knockout samples were subjected to SDS-PAGE. ab157194 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-SPR antibody [EPR9290] (ab157194)

All lanes : Anti-SPR antibody [EPR9290] (ab157194) at 1/1000 dilution

Lane 1 : A549 cell lysate
Lane 2 : MCF7 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 28 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

ab157194 showing +ve staining in Human normal pancreas.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

ab157194 showing +ve staining in Human heart.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

ab157194 showing +ve staining in Human endometrial carcinoma.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

ab157194 showing +ve staining in Human colonic adenocarcinoma.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-SPR antibody [EPR9290] (ab157194)

Intracellular Intracellular Flow Cytometic analysis of permeabilized MCF7 cells labeling TPST2 with ab157194 at 1/100 (red) or a rabbit IgG (negative) (green).
Immunocytochemistry/ Immunofluorescence - Anti-SPR antibody [EPR9290] (ab157194)

Immunofluorescent analysis of HeLa cells labeling TPST2 with ab157194 at 1/100 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

Immunohistochemical analysis of paraffin-eEmbedded Human thyroid gland carcinoma tissue labeling TPST2 with ab157194 at 1/100 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunoprecipitation - Anti-SPR antibody [EPR9290] (ab157194)

Detection of TPST2 by Western Blot of Immunprecipitate. MCF7 cell lysate immunoprecipitated using ab157194 at 1/10 dilution.
Anti-SPR antibody [EPR9290] (ab157194)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab157194? Please let us know so that we can cite the reference in this datasheet.

ab157194 has been referenced in 3 publications.

Wu Y et al. Sepiapterin reductase promotes hepatocellular carcinoma progression via FoxO3a/Bim signaling in a nonenzymatic manner. Cell Death Dis 11:248 (2020). PubMed: 32312975
Mooney MR et al. Anti-tumor effect of sulfasalazine in neuroblastoma. Biochem Pharmacol 162:237-249 (2019). PubMed: 30639262
Sallin O et al. Semisynthetic biosensors for mapping cellular concentrations of nicotinamide adenine dinucleotides. Elife 7:N/A (2018). PubMed: 29809136

Customer reviews and Q&As

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Application

Western blot

Sample

Rabbit Tissue lysate - whole (brain)

Gel Running Conditions

Non-reduced Denaturing

Loading amount

0.2 µg

Treatment

hypoxic 40 minutes at E25

Specification

brain

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 25 2018

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (3)

Customer reviews and Q&As

Western blot abreview for Anti-SPR antibody [EPR9290]

Post-translational
modifications