BiP | lumenal-binding protein

345 €

AS09 614 | clonality: polyclonal | host: hen | reactivity: Arabidopsis thaliana, Hordeum vulgare, Spinacia oleracea, Zea mays

PRODUCT INFORMATION IN PDF

product information

background

BiP2 (Binding immunoglobulin protein) is localized in endoplasmic reticulum lumen (ER) and plays a role in protein assembly inside ER. BiP protein is abundant under all growth conditions but its synthesis can increase under conditions that lead to the accumulation of unfolded polypeptides in endoplasmic reticulum (ER). Alternative name: AtBP2

immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana BiP proteins: BiP1 At5g28540 Q9LKR3, BiP2 At5g42020 F4K007 , BiP3 At1g09080 Q8H1B3

antibody format

hen

polyclonal,

affinity purified IgY in PBS pH 7.4, liquid

quantity

100 µg

storage

store at 4°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

tested applications

western blot (WB), immunofluorescence (IF)

related products

AS09 481 | BiP2 | lumenal-binding protein 2, rabbit antibody

AS09 615 | BiP2 | lumenal-binding protein 2, goat antibody

antibodies to plant endomembrane system proteins

additional information

Antibody solution contains 0.02% sodium azide as preservative

Method for plant ER isolation is available here.

application information
recommended dilution		1: 2000 with standard ECL (WB), 1: 1000 (IF)
expected \| apparent MW		73.5 \| 80 kDa
confirmed reactivity		dicots including: Arabidopsis thaliana, Spinacia oleracea, monocots including: Hordeum vulgare, Zea mays
predicted reactivity		dicots including: Nicotiana tabacum, Spinacia oleracea, monocots: Oryza sativa, Zea mays, trees: Picea sitchensis, Populus trichocarpa, moss: Physcomitrella patens
not reactive in		no confirmed exceptions from predicted reactivity known in the moment
additional information		Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
selected references		to be added when available, antibody released in September 2010

application example

5 µg of total protein from A.thaliana (1), H. vulgare (2), Z.mays (3), S. oleracea (4), extracted with Agrisera PEB extraction buffer (AS08 300) were separated on 4-12% SDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-hen IgY horse radish peroxidase conjugated, from Agrisera AS09 603) diluted to 1:50 000 for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL detection reagent according to the manufacturers instructions. Exposure time was 5 seconds.

western blot detection of plant BiP using hen antibody

immunofluorescence

immunolocalization of plant BiP using hen antibody

BiP localization in 5 days old Arabidopsis thaliana roots. BiP signal shown in green, DAPI in blue. The material has been fixed in para-formaldehyde for 30 minutes. Tissue cleaning has been performed before immunolocalization. Chicken anti-BiP primary antibody was diluted in 1: 1000 and DyLight®488 conjugated goat anti-chicken secondary antibody AS09 622 (green color) was diluted in 1: 1000. Co-staining with DAPI visualized nucleus (blue color). Scale bar – 10 µm.

Courtesy Dr. Taras Pasternak, Freiburg University

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