SPINK1 Antibody (839305) [Unconjugated]

Western blot shows lysates of human pancreas tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human SPINK1 Monoclonal Antibody (Catalog # MAB7496) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for SPINK1 at approximately 6 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of human pancreas tissue, loaded at 0.5 mg/mL. A specific band was detected for SPINK1 at approximately 6 kDa (as indicated) using 100 µg/mL of Mouse Anti-Human SPINK1 Monoclonal Antibody (Catalog # MAB7496). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Androgen signaling negatively regulates SPINK1 expression in prostate cancer.a Immunoblot for SPINK1 in 22RV1 cells stimulated with R1881 (10 nM) (top). QPCR data showing relative expression of SPINK1&KLK3 in the same cells (bottom). Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reprogramming factor SOX2&AR transcriptional co-repressor REST modulate SPINK1 expression.a Schematic showing SOX2 binding elements (S1, S2,&S3) on the SPINK1 promoter (top). ChIP-qPCR data for SOX2 occupancy on the SPINK1 promoter in wildtype LNCaP&LNCaP-AI cells (androgen-deprived for 15 days) (bottom). b Same as in a, except 22RV1 cells. c ChIP-qPCR data for RNA Pol-II binding on the SPINK1 promoter using cells as a. d Same as c, except 22RV1 cells. e Immunoblot for SOX2&SPINK1 in siRNA mediated SOX2-silenced LNCaP-AI&control cells. f Same as e, except 22RV1 cells.Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Androgen signaling negatively regulates SPINK1 expression in prostate cancer. h Immunoblot showing SPINK1 expression in VCaP cells treated with enzalutamide (top). QPCR data showing relative expression of SPINK1, KLK3,&ERG (bottom). Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reprogramming factor SOX2&AR transcriptional co-repressor REST modulate SPINK1 expression.a Schematic showing SOX2 binding elements (S1, S2,&S3) on the SPINK1 promoter (top). ChIP-qPCR data for SOX2 occupancy on the SPINK1 promoter in wildtype LNCaP&LNCaP-AI cells (androgen-deprived for 15 days) (bottom). b Same as in a, except 22RV1 cells. c ChIP-qPCR data for RNA Pol-II binding on the SPINK1 promoter using cells as a. d Same as c, except 22RV1 cells. e Immunoblot for SOX2&SPINK1 in siRNA mediated SOX2-silenced LNCaP-AI&control cells. f Same as e, except 22RV1 cells. g QPCR data showing relative expression of SOX2&SPINK1 upon transient SOX2 overexpression in LNCaP cells (left). Immunoblot for SOX2&SPINK1 expression(right). Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Androgen signaling negatively regulates SPINK1 expression in prostate cancer. l Immunoblot showing SPINK1 expression in 22RV1 cells as indicated in k (top). QPCR data showing relative expression of SPINK1&KLK3 using same cells in k (bottom)Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Androgen signaling negatively regulates SPINK1 expression in prostate cancer.a Immunoblot for SPINK1 in 22RV1 cells stimulated with R1881 (10 nM) (top). QPCR data showing relative expression of SPINK1&KLK3 in the same cells (bottom). b Immunostaining for SPINK1&AR in 22RV1 cells stimulated with R1881 (10 nM). c Same as b, except dot plot represents quantification for SPINK1 mean fluorescence intensity (MFI) per unit area shown as arbitrary units (AU). d Same as a, except VCaP cells used. Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reprogramming factor SOX2&AR transcriptional co-repressor REST modulate SPINK1 expression.a Schematic showing SOX2 binding elements (S1, S2,&S3) on the SPINK1 promoter (top). ChIP-qPCR data for SOX2 occupancy on the SPINK1 promoter in wildtype LNCaP&LNCaP-AI cells (androgen-deprived for 15 days) (bottom). b Same as in a, except 22RV1 cells. c ChIP-qPCR data for RNA Pol-II binding on the SPINK1 promoter using cells as a. d Same as c, except 22RV1 cells. e Immunoblot for SOX2&SPINK1 in siRNA mediated SOX2-silenced LNCaP-AI&control cells. Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reprogramming factor SOX2&AR transcriptional co-repressor REST modulate SPINK1 expression.k Immunoblot for the REST&SPINK1 levels in 22RV1 cells treated with Casein Kinase 1 inhibitor (iCK1) as indicated (top). QPCR data for relative SPINK1&SYP expression (bottom). Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

SPINK1 promotes EMT, stemness&chemoresistance in prostate cancer.a Immunoblot analysis for SPINK1, E-Cadherin&Vimentin levels in stable SPINK1-silenced (shSPINK1-1&shSPINK1-2)&control (shSCRM) 22RV1 cells. Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western
• Clone: 839305
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

SPINK1 Antibody (839305) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human SPINK1
  Asp24-Cys79
  Accession # P00995
• Specificity: Detects human SPINK1 in ELISAs.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: SPINK1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Simple Western 100 ug/mL
  • Western Blot 2 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 µm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for SPINK1 Antibody (839305) [Unconjugated]

• pancreatic secretory trypsin inhibitor
• PCTT
• PCTTSpink3
• PSTI
• PSTISerine protease inhibitor Kazal-type 1
• serine peptidase inhibitor, Kazal type 1
• serine protease inhibitor, Kazal type 1
• SPINK1
• Spink3
• TATI
• TATITumor-associated trypsin inhibitor

Background

SPINK1 (Serine Protease Inhibitor Kazal-type 1; also TATI and PST1) is a 6-7 kDa secreted polypeptide initially identified as a tumor-derived trypsin inhibitor. It is widely expressed and found in cells diverse as pancreatic acinar cells, columnar cells of the stomach, renal collecting duct epithelium, and ureteric transitional plus breast epithelium. SPINK1 is known to be secreted with pancreatic zymogens, and apparently inactivates prematurely-activated trypsin, thus protecting the pancreas from trypsin-mediated enzyme activation. It also is reported to regulate cell migration and proliferation, the latter effect attributed to its structural resemblance to EGF and its ability to bind to activate the EGFR. Mature human SPINK1 is 56 amino acids (aa) in length (aa 24-79). It contains one Kazal-like domain (aa 26-79) that possesses a potential proteolytic cleavage site between Lys41-Ile42. An 11-12 kDa form in SDS-PAGE has been reported for SPINK1, possibly reflecting dimerization. Mature human SPINK1 shares 66% aa sequence identity with mouse Spink3, the mouse equivalent to human SPINK1.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for SPINK1 Antibody (MAB7496)(5)

Publications using MAB7496

• Hu WY, Hu DP, Xie L et al. Keratin Profiling by Single-Cell RNA-Sequencing Identifies Human Prostate Stem Cell Lineage Hierarchy and Cancer Stem-Like Cells International Journal of Molecular Sciences 7/28/2021 [PMID: 34360875]
• Hu WY, Hu DP, Xie L et al. Keratin Profiling by Single-Cell RNA-Sequencing Identifies Human Prostate Stem Cell Lineage Hierarchy and Cancer Stem-Like Cells International Journal of Molecular Sciences 2021-07-28 [PMID: 34360875]
• R Tiwari, N Manzar, V Bhatia, A Yadav, MA Nengroo, D Datta, S Carskadon, N Gupta, M Sigouros, F Khani, M Poutanen, A Zoubeidi, H Beltran, N Palanisamy, B Ateeq Androgen deprivation upregulates SPINK1 expression and potentiates cellular plasticity in prostate cancer Nat Commun, 2020-01-20;11(1):384. 2020-01-20 [PMID: 31959826] (Western Blot, Human)
• Shipra Shukla, Joanna Cyrta, Devan A. Murphy, Edward G. Walczak, Leili Ran, Praveen Agrawal et al. Aberrant Activation of a Gastrointestinal Transcriptional Circuit in Prostate Cancer Mediates Castration Resistance Cancer Cell 12/11/2017 [PMID: 29153843]
• Shipra Shukla, Joanna Cyrta, Devan A. Murphy, Edward G. Walczak, Leili Ran, Praveen Agrawal et al. Aberrant Activation of a Gastrointestinal Transcriptional Circuit in Prostate Cancer Mediates Castration Resistance Cancer Cell 2017-12-11 [PMID: 29153843] (Western Blot, Human)

Bioinformatics

• Gene Symbol: SPINK1
• Uniprot:
  • P00995()