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PIP2;2 | Plasma membrane aquaporin 2b

AS09 490 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Raphanus sativus

PIP2;2 | Plasma membrane aquaporin 2b in the group Antibodies Plant/Algal  / Membrane Transport System / Plasma membrane at Agrisera AB (Antibodies for research) (AS09 490)
PIP2;2 | Plasma membrane aquaporin 2b



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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from N-terminal part of Raphanus sativus PAQ2b UniProt: Q9MB99

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 100 ĩl
Reconstitution For reconstitution add 100 ĩl of sterile water
Storage Store lyophilized/reconstituted at -20°C; make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications ELISA (ELISA), Western blot (WB)
Recommended dilution 1 : 8000 (ELISA), 1 : 2000 (WB)
Expected | apparent MW

30 | 28 kDa (Raphanus sativus)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Fragaria ananassa, Raphanus sativus
Predicted reactivity Brassica sp.
Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples

Application example

western blot using anti-PIP2;2 antibodies

1 µg and 10 µg of crude membrane fraction/lane from Arabidopsis thaliana  were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-PIP2;2 antibodies (AS09 490, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

PIP proteins usually show a faint band of dimeric form at 55 kDa addition to the major monomer band of 28 kDa even in the presence of SDS.

Additional information

Additional information

0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.

Antibodies will detect target protein in a few  µg of a crude preparation loaded per well. If purified preparations of vacuolar and plasma membranes are used, one µg load per well should be sufficient.

Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.

Antiboy will weakly react with PIP2;1 and theoretically with PIP2;3, however this protein is very scarcely expressed in plant tissues. The mRNA content is 1 % of that for PIP2;2, therefore reactivity to PIP2;3 was neglegible in our experiments.

Triton X-100 should not be included in the protein extraction buffer, when cell organelles or membrane proteins must be separated from soluble proteins. Because, Triton X breaks membrane structure and solubilizes most membranes proteins. Furthermore, it should be noted that Triton X at high concentrations binds SDS and mask the detergent effect of SDS for SDS-PAGE. Also, micelles of Triton X behave as a large complex with molecular mass of 90 kDa at high concentrations in SDS-PAGE.

Related products

Background

Background

PIP2;2 is a plasma membrane aquaporin.

Product citations

Selected references Brillada et al. (2020). Exocyst subunit Exo70B2 is linked to immune signalling and autophagy, The Plant Cell, 2020;, koaa022, https://doi.org/10.1093/plcell/koaa022
immunogen:

KLH-conjugated synthetic peptide derived from N-terminal part of Raphanus sativus PAQ2b UniProt: Q9MB99

Reconstitution: For reconstitution add 100 ĩl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 100 ĩl
storage: Store lyophilized/reconstituted at -20°C; make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications: ELISA (ELISA), Western blot (WB)
recommended dilution: 1 : 8000 (ELISA), 1 : 2000 (WB)
calculated | apparent molecular mass [kDa]:

30 | 28 kDa (Raphanus sativus)

Confirmed reactivity: Arabidopsis thaliana, Fragaria ananassa, Raphanus sativus
predicted reactivity: Brassica sp.
Species of your interest not listed? Contact us
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer):

Application example

western blot using anti-PIP2;2 antibodies

1 µg and 10 µg of crude membrane fraction/lane from Arabidopsis thaliana  were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-PIP2;2 antibodies (AS09 490, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

PIP proteins usually show a faint band of dimeric form at 55 kDa addition to the major monomer band of 28 kDa even in the presence of SDS.

additional information:

0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.

Antibodies will detect target protein in a few  µg of a crude preparation loaded per well. If purified preparations of vacuolar and plasma membranes are used, one µg load per well should be sufficient.

additional information (application):

Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.

Antiboy will weakly react with PIP2;1 and theoretically with PIP2;3, however this protein is very scarcely expressed in plant tissues. The mRNA content is 1 % of that for PIP2;2, therefore reactivity to PIP2;3 was neglegible in our experiments.

Triton X-100 should not be included in the protein extraction buffer, when cell organelles or membrane proteins must be separated from soluble proteins. Because, Triton X breaks membrane structure and solubilizes most membranes proteins. Furthermore, it should be noted that Triton X at high concentrations binds SDS and mask the detergent effect of SDS for SDS-PAGE. Also, micelles of Triton X behave as a large complex with molecular mass of 90 kDa at high concentrations in SDS-PAGE.

background:

PIP2;2 is a plasma membrane aquaporin.

All references: Brillada et al. (2020). Exocyst subunit Exo70B2 is linked to immune signalling and autophagy, The Plant Cell, 2020;, koaa022, https://doi.org/10.1093/plcell/koaa022

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