BiP | Lumenal-binding protein (rabbit antibody)

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

73.5 | 80 kDa

5 µg of total protein from A.thaliana (1), H. vulgare (2), P. sativum (3)*, Z. mays (4), C. sativus(5), S. tuberosum (6), S. oleracea (7), S. lycopersicum (8) P. patens (9)*, C. reinhardtii (10)  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 5 % non-fat milk in TBS-T,  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-rabbit IgG horse radish peroxidase conjugated, from  Agrisera AS09 602) 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 of extreme femtogram range, according to the manufacturers instructions.  Exposure time was 5 seconds. * Lack of the signal or its low signal intensity in those samples can be due to the sample biology. If you work with those species, please inquire.

Immunolocalization

Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel. This antibody has so far not worked in IP.

AS09 615 | Anti-BiP2 | lumenal-binding protein 2, goat antibodies
AS09 614 | Anti-BiP2 | lumenal-binding protein 2, chicken antibodies
AS09 481PRE | BiP | lumenal-binding protein, pre-immune serum
antibodies to plant endomembrane system proteins

Plant protein extraction buffer

Secondary antibodies

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).