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

signalword

Danger

hazcat

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

storage_class_code

11 - Combustible Solids

WGK Germany

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

Wilson, M.B. and Nakane, P.K. et al.
Immunofluorescence and Related Staining Techniques, 215-215 (1978)
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...
Jianghong Yan et al.
Biochemistry and molecular biology education : a bimonthly publication of the International Union of Biochemistry and Molecular Biology, 46(4), 373-381 (2018-07-11)
Medical laboratory technology major was set up to meet rapid development of science and medical research technology in 2013. Students majoring in medical laboratory had learnt a lot of techniques distributed among different specialized courses. But, they did not understand...
Jia Yang et al.
Journal of plant physiology, 226, 1-11 (2018-04-25)
Stress-adapted wild plants are natural sources of novel genes for molecular breeding. Here, we conducted a transcriptional analysis of Pinus sylvestris var. mongolica Litv, an evergreen pine in northeastern China, to identify a novel CALMODULIN-LIKE protein-encoding gene, PsCML1, no significant...

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