All Photos(1)

L6397

Sigma-Aldrich

Concanavalin A from Canavalia ensiformis (Jack bean)

peroxidase conjugate, lyophilized powder

Synonym(s):
ConA
NACRES:
NA.32

Quality Level

biological source

Canavalia ensiformis (jack bean)

form

lyophilized powder

application(s)

blood typing: suitable
fractionation: suitable

impurities

carbohydrate, essentially free

conjugate

peroxidase conjugate

storage temp.

−20°C

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Application

General Western Blot Protocol:
  • Glycoprotein sample size: 500ng
  • Lectin Concentration: 0.1ug/ml

  1. Load samples at 500 ng of glycoprotein per lane
  2. Run 4-20% Bis-Tris SDS page gel
  3. Transfer gel to a PVDF membrane
  4. Block membrane for 1 hr at RT with RIPA buffer (R0278 Sigma)
  5. Incubate HRP lectin at 0.1ug/ml with RIPA buffer for 2 hours at RT
  6. Wash membrane 5 x 5 minutes with 25ml RIPA buffer
  7. Detect using chemiluminescent substrate (CPS1-120)
Detects glycoproteins containing α-D-mannose, α-D-glucose when used with appropriate peroxidase substrate

Biochem/physiol Actions

Con A is not blood group specific but has an affinity for terminal α-D-mannosyl and α-D-glucosyl residues. Ca2+ and Mn2+ ions are required for activity. Con A dissociates into dimers at pH 5.6 or below. Between pH 5.8 and pH 7.0, Con A exists as a tetramer; above pH 7.0 higher aggregates are formed. Con A exhibits mitogenic activity which is dependent on its degree of aggregation. Succinylation results in an active dimeric form which remains a dimer above pH 5.6.

Packaging

Package size based on protein content

Unit Definition

One unit will form 1 mg purpurogallin in 20 sec from pyrogallol at pH 6.0 at 20 °C.

Physical form

Lyophilized powder containing tris-citrate buffer salts and trace calcium and manganese

Preparation Note

Prepared from peroxidase type VI (P8375), using the method of Wilson and Nakane, which promotes conjugation but prevents the interaction between Con A and peroxidase sugar residues.

Analysis Note

Where reported, agglutination activity is expressed in μg/ml and is determined from serial dilutions in phosphate buffered saline, pH 6.8, containing Ca2+ and Mn2+ of a 1 mg per mL solution. This activity is the lowest concentration to agglutinate a 2% suspension of human erythrocytes after 1 hr incubation at 25 °C.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Hazard Classifications

Repr. 2 - Resp. Sens. 1 - Skin Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US),Eyeshields,Gloves

Certificate of Analysis

Certificate of Origin

Johanna Kabbert et al.
The Journal of experimental medicine, 217(11) (2020-07-09)
The gut is home to the body's largest population of plasma cells. In healthy individuals, IgA is the dominating isotype, whereas patients with inflammatory bowel disease also produce high concentrations of IgG. In the gut lumen, secretory IgA binds pathogens...
Frederik Bühring Bjørkskov et al.
Scientific reports, 7(1), 16899-16899 (2017-12-06)
The sparse number of high-resolution human membrane protein structures severely restricts our comprehension of molecular physiology and ability to exploit rational drug design. In the search for a standardized, cheap and easily handled human membrane protein production platform, we thoroughly...
Wilson, M.B. and Nakane, P.K. et al.
Immunofluorescence and Related Staining Techniques, 215-215 (1978)
Lingyan Li et al.
FEBS letters, 594(4), 665-677 (2019-10-31)
Adaptation to higher temperatures would increase the environmental competitiveness of psychrophiles, organisms that thrive in low-temperature environments. Methanolobus psychrophilus, a cold wetland methanogen, 'evolved' as a mesophile, growing optimally at 30 °C after subculturings, and cells grown with ample substrates exhibited...
Wilhelm Bohr et al.
PloS one, 5(12), e16000-e16000 (2011-01-07)
The CCN family of proteins, especially its prominent member, the Connective tissue growth factor (CTGF/CCN2) has been identified as a possible biomarker for the diagnosis of fibrotic diseases. As a downstream mediator of TGF-β1 signalling, it is involved in tissue...

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