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ELISA Synblock

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Synblock solution (BUF034) used for blocking non-specific binding sites in a Mincle binding assay
Image caption:
T. forsythia S-layer binds Mincle.
(A) Binding of S-layer to Mincle. T. forsythia S-layer immobilized in microtiter plate wells was incubated with Mincle-Fc fusion protein or MMR-Fc in lectin binding buffer (TBS, 5 mM CaCl2). As control, ManLAM was immobilized in wells. After washing wells with buffer, binding was detected with HRP-coupled goat anti-human IgG-Fc and chromogenic substrate TMB. Color was read by measuring absorbance at 450 nm. As shown, S-layer bound Mincle in a dose dependent manner and MMR as control bound ManLAM but not S-layer. (B) Mincle binding to T. forsythia S-layer is Ca2+-dependent. To analyze the Ca2+ dependency of Mincle binding to S-layer, Mincle-Fc was incubated with plate bound S-layer in Ca2+ (TBS, 5 mM CaCl2) or EDTA containing buffer (TBS, 10 mM EDTA). Plate bound TDB was used a control ligand or Mincle binding. Binding was then detected as above. (C) S-layer dose dependently inhibited binding of Mincle to TDB. Plate bound TDB was incubated with Mincle-Fc in lectin binding buffer with increasing concentrations of S-layer. Wells were washed with elution buffer and binding was measured as above. Each value represents the mean (± SD) of 3 values measured in one representative assay. Data are representative of three independent experiments with similar results; *, P<0.05.

From: Chinthamani S, Settem RP, Honma K, Kay JG, Sharma A (2017)
Macrophage inducible C-type lectin (Mincle) recognizes glycosylated surface (S)-layer of the periodontal pathogen Tannerella forsythia.
PLoS ONE 12(3): e0173394.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.

ELISA Synblock

Product Type
Accessory Reagent
Specificity
ELISA Synblock

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Product Code Applications Pack Size List Price Your Price Qty
BUF034A
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
E 100 ml loader
List Price Your Price
loader
BUF034B
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
E 500 ml loader
List Price Your Price
loader
BUF034C
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
E 1000 ml loader
List Price Your Price
loader

ELISA Synblock is an ELISA buffer designed to avoid false positive results associated with buffers containing animal proteins (e.g. BSA) and reduce non-specific background signals in ELISA assays without the addition of the usual protein additives.

Intended Use
BUF034A, BUF034B: ELISA SynBlock is a novel protein-free blocking buffer suitable for use in all ELISA formats requiring maximum blocking strength. With Tween and synthetic blocking agents, the inert nature of this unique buffer, enables maximum reduction of non-specific binding and interference associated particularly with sandwich ELISA assays.

Additional molecular stabilizers and an antimicrobial agent provide a long-term stable environment for coating antigen or capture antibody. Plates can be blocked at room temperature and stored once dried for up to a year at +4°C.

N.B. SYNBLOCK is not suitable for use on Immunolon-2 plates. Bio-Rad recommends the use of BUF033A for this purpose.
BUF034C: ELISA SynBlock is a novel protein-free blocking buffer suitable for use in all ELISA formats requiring maximum blocking strength. With detergent and synthetic blocking agents, the inert nature of this unique buffer enables maximum reduction of non-specific binding and interference associated particularly with sandwich ELISA assays.

Additional molecular stabilizers and an antimicrobial agent provide a long-term stable environment for coating antigen or capture antibody. Plates can be blocked at room temperature and stored once dried for up to a year at +4°C.

N.B. SynBlock is not suitable for use on Immunlon® 2 plates. Bio-Rad recommends the use of BUF033A for this purpose.
Product Form
Ready to use - liquid
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
BUF034A: <0.1% sodium azide (NaN3)
BUF034B, BUF034C:
0.05%Sodium Azide
Regulatory
For research purposes only
Guarantee
Guaranteed until date of expiry. Please see product label.

Store at +4oC.

DO NOT FREEZE

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
ELISA Neat
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Instructions For Use
1. Coat ELISA plate with antibody or antigen as required.

2. After incubation, remove the coating solution and wash the plate x2 with wash buffer. BUF031A can be used for this purpose.

3. Add 300-400ul of BUF034A and incubate for 2-24 hours. Use a volume equal to or greater than the volume of coating solution.

4. After removal of the blocking buffer continue with the assay or dry the plate for long-term storage at +4°C.
1. Coat ELISA plate with antibody or antigen as required.

2. After incubation, remove the coating solution and wash the plate x2 with wash buffer. BUF031A can be used for this purpose.

3. Add 300-400ul of BUF034B and incubate for 2-24 hours. Use a volume equal to or greater than the volume of coating solution.

4. After removal of the blocking buffer continue with the assay or dry the plate for long-term storage at +4°C.
1. Coat ELISA plate with antibody or antigen as required.

2. After incubation, remove the coating solution and wash the plate x2 with wash buffer. BUF031A can be used for this purpose.

3. Add 300-400ul of BUF034C and incubate for 2-24 hours. Use a volume equal to or greater than the volume of coating solution.

4. After removal of the blocking buffer continue with the assay or dry the plate for long-term storage at +4°C.

Description Product Code Applications Pack Size List Price Your Price Quantity
5x ELISA Coating Buffer BUF030A E 100 ml
List Price Your Price
Description 5x ELISA Coating Buffer
10x ELISA Wash Buffer BUF031A E 100 ml
List Price Your Price
Description 10x ELISA Wash Buffer

References for ELISA Synblock

  1. Afrough, B. et al. (2007) Identification and elimination of false-positives in an ELISA-based system for qualitative assessment of glycoconjugate binding using a selection of plant lectins.
    Biotechniques. 43 (4): 458, 460, 462 passim.
  2. Dalley, D. et al. (2008) Development and evaluation of a gamma-interferon assay for tuberculosis in badgers (Meles meles).
    Tuberculosis (Edinb). 88: 235-43.
  3. Ahmed, R.R. et al. (2010) BACE1 and BACE2 enzymatic activities in Alzheimer's disease.
    J Neurochem. 112: 1045-53.
  4. Chambers, M.A. et al. (2009) Performance of TB immunodiagnostic tests in Eurasian badgers (Meles meles) of different ages and the influence of duration of infection on serological sensitivity.
    BMC Vet Res. 5: 42.
  5. Thompson, R. et al. (2011) Optimization of the enzyme-linked lectin assay for enhanced glycoprotein and glycoconjugate analysis.
    Anal Biochem. 413: 114-22.
  6. Kuramitz, H. et al. (2012) Multiplexed assay for proteins based on sequestration electrochemistry using the protein binding electroactive magnetic microbeads.
    Anal Sci. 28 (1): 77.
  7. Dwek, M.V. et al. (2010) A sensitive assay to measure biomarker glycosylation demonstrates increased fucosylation of prostate specific antigen (PSA) in patients with prostate cancer compared with benign prostatic hyperplasia.
    Clin Chim Acta. 411 (23-24): 1935-9.
  8. Verhelst, R. et al. (2010) The effects of plant polyphenols on enterotoxigenic Escherichia coli adhesion and toxin binding
    Livestock Science. 133 (1-3): 101-3
    9. View The Latest Product References
  9. Verhelst, R. et al. (2013) E. coli heat labile toxin (LT) inactivation by specific polyphenols is aggregation dependent.
    Vet Microbiol. 163 (3-4): 319-24.
  10. Greenwell P et al. (2008) Purification and analysis of DNases of Tritrichomonas foetus: evidence that these enzymes are glycoproteins.
    Int J Parasitol. 38 (7): 749-56.
  11. Beckett, T.L. et al. (2013) A ketogenic diet improves motor performance but does not affect β-amyloid levels in a mouse model of Alzheimer's disease.
    Brain Res. 1505: 61-7.
  12. Abdul, H.M. et al. (2009) Cognitive decline in Alzheimer's disease is associated with selective changes in calcineurin/NFAT signaling.
    J Neurosci. 29 (41): 12957-69.
  13. Niedowicz DM et al. (2013) Leptin regulates amyloid β production via the γ-secretase complex.
    Biochim Biophys Acta. 1832 (3): 439-44.
  14. Martin SB et al. (2014) Synaptophysin and synaptojanin-1 in Down syndrome are differentially affected by Alzheimer's disease.
    J Alzheimers Dis. 42 (3): 767-75.
  15. Wilcock, D.M. et al. (2015) Down syndrome individuals with Alzheimer's disease have a distinct neuroinflammatory phenotype compared to sporadic Alzheimer's disease.
    Neurobiol Aging. 36 (9): 2468-74.
  16. Chinthamani S et al. (2017) Macrophage inducible C-type lectin (Mincle) recognizes glycosylated surface (S)-layer of the periodontal pathogen Tannerella forsythia.
    PLoS One. 12 (3): e0173394.
  17. LeVine, H. 3rd.et al. (2017) Down syndrome: age-dependence of PiB binding in postmortem frontal cortex across the lifespan.
    Neurobiol Aging. 54: 163-9.

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