Anti-PDI antibody [RL90] - ER Marker (ab2792)

human and mouse platelets - used to work before very nicely
now: no bands in WB with lot GR52326-2, lot GR64719-1 also gave no bands
Ab: 1/2000, 1/200
block: 5% milk
secondary: OK

other PDI antibodies work well

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx for ab5484.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

This antibody is staining non specifically in ICC.

ANSWER:

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab2792.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Dear Tanya,

thank you very much for the reply.

Could you therefore please send us ab5484 as replacement? I will then also recommend the customer to use more protein!

Further, we will then also send you the generated data!

Thank you in advance!

Best regards,

ANSWER:

Thank you for your response.

This is to let you know that I have placed a new order for you - for one vial of ab2792 as a free of charge replacement and the new order number is 11140.

I hope the second vial will work as it is expected, and please do let me know how you are getting on with this product.

I have a technical enquiry regarding the immunogen used for the production of the PDI antibody [RL90]. I have used the antibody and find that it works well, detecting a ~57kDa protein by immunoblot. However, there are several PDI proteins of ~57kDa and the datasheet is not clear which of these was used as the immunogen - my explanation below is quite detailed, but hopefully it will help you answer my question...

The immunogen is stated as "Full length native protein (purified) (Rat)".

Some background: The PDI family comprises over 10 different proteins, with 'PDI' itself being the prototypical member of this family, and the various other PDI family members being given various different names. The terminology is confused, and confusing, between the different proteins, with most of them having at least several names.

'PDI' itself is also known as PDIA1 or P4HB (human Entrez Gene ID 5034).

A second family member, which is also sometimes confusingly called simply 'PDI', is PDIA2, or PDIp, encoded by a different gene (human Entrez Gene ID 64714).

A third protein, encoded by yet another gene, is PDIA3 (more commonly called ERp57 or GRP58, human Entrez Gene ID 2923).

The details listed on the RL90 antibody product page variously point towards database entries for either PDIA1, PDIA2 or even PDIA3. Some of these links on your site are:

Entrez 64714 - human PDIA2 Entrez 14827 - mouse PDIA3 Entrez 25506 - rat P4HB (PDIA1) Genecard GC16P000274 - human PDIA2 Omim 176790 - human P4HB (PDIA1) SwissProt P07237 - human PDIA1 SwissProt P09103 - mouse PDIA1 SwissProt P04785 - rat PDIA1 Unigene 410578 - human (no longer available) Unigene 4234 - rat P4HB (PDIA1)

In addition, the 'Alternative names' page lists the names for both PDIA1 (P4HB) and PDIA2 (PDIp) as if they are synonyms, which they are not.

Could you please tell me which one of the PDI family members was used as the immunogen for this antibody? (And secondarily, it may be useful for other customers to have the datasheet for this antibody updated to reflect only the PDI protein that was used as the immunogen).

Thanks very much for your time.

ANSWER:

Thank you for your email.

We have checked with the source of the antibody through their development records for an answer to this question, but we were unsuccessful - it just says PDI rather than a specific isoform. The original reference for this antibody is as follows:

C.S. Kaetzel et al. Protein Disulphide-Isomerase from Human Placenta & Rat Liver; Purification & Immunological Characterization with Monoclonal Antibodies. J. Biol. Chem., 241: 39-47, 1987.

We have scanned the materials and methods section of this paper which describes isolation of the PDI protein and attached it to this email for reference. It does not describe an isoform, rather the PDI was isolated by size. The paper was published probably before the different isoforms became known.

I'm sorry I can't be of more help.

I am not seeing any bands. I am using the ab at 1/1000 on mouse liver extracts between 50 to 150 ug.

I am using an anti-IgG1 secondary.

ANSWER:

Thank you for your enquiry.

As we discussed, you should be using an anti-IgG2a secondary antibody. Please try this and if it does not work then please let me know.

Here is the isotype: Isotype IgG2a.

Here also is the datasheet:

PDI antibody [RL90] - ER Marker (ab2792) datasheet Datasheet Abreviews™ (2) Enquiries (8) Protocols Related products

printer friendly datasheet · Sodium Azide MSDS Immunocytochemistry - PDI antibody [RL90] - ER Marker (ab2792)

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Immunocytochemistry - PDI antibody [RL90] - ER Marker (ab2792)

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Availability If you place an order for a unit of ab2792 now (9:31 AM EST on 10 April 2006) you should get it on Tuesday 11 April 2006 Select country Argentina Australia Austria Bahrain Bangladesh Belarus Belgium Brazil Bulgaria Canada Chile China Colombia Croatia Cyprus Czech Republic Denmark Egypt Estonia Finland France Germany Greece Guam Hong Kong Hungary Iceland India Indonesia Iran Ireland Israel Italy Jamaica Japan Jordan Korea, Republic of Kuwait Latvia Lebanon Lithuania Luxembourg Malaysia Malta Mexico Monaco Nepal Netherlands New Zealand Nigeria Norway Pakistan Panama Poland Portugal Puerto Rico Reunion Romania Russian federation Saudi Arabia Senegal Singapore Slovakia Slovenia South Africa Spain Sri lanka Sweden Switzerland Taiwan Thailand Trinidad & Tob Tunisia Turkey U.A.E Ukraine United Kingdom United States Uruguay Venezuela Vietnam Yugoslavia Price Code Size Price (USD) Points * ab2792 100 µl $ 299.00 19 We accept credit cards and purchase order numbers. You can also order by email, fax, phone and post. (details) Freight/Packing $ 28.00 (for United States) Abpoints * Get discounts / gifts by earning Abpoints Earn 19 Abpoints by placing your order online Earn 50 Abpoints by reviewing this product Read about Abpoints

-------------------------------------------------------------------------------- Product Name PDI antibody [RL90] - ER Marker (read about markers) See all PDI antibodies (5)... Product type Primary antibodies Description Mouse monoclonal [RL90] to PDI - ER Marker Immunogen Full length native protein (purified) (Rat). Species Reactivity Reacts with Human, Mouse, Rat, Dog (see review) and Hamster. Not yet tested in other species. Tested applications (see all reviews) Western blot, Immunoprecipitation, Immunohistochemistry (Formalin-fixed paraffin-embedded sections), Immunohistochemistry (Frozen sections), Immunofluorescence, Immunocytochemistry reviews Application notes (see key) Recommended dilutions Immunofluorescence: 1/100 Immunohistochemistry: 1/100 Western Blot: 1/1000 Immunoprecipitation: Assay dependent. Not tested in other applications.

Optimal dilutions / concentrations should be determined by the end user.

By Western blot, this antibody detects a protein at 59 kDa representing PDI from rat liver extract or a slightly higher protein at 61 kDa representing PDI from human liver extract.

Immunohistochemical staining of PDI in rat intestine yields a pattern consistent with cytoplasmic staining.

In immunoprecipitation procedures, this antibody has been shown to inhibit the activity of PDI in vitro.

This antibody has also been found to inhibit disulfide bond reduction of the HIV protein, gp120, at the cell surface of CHO cells and human lymphoid cells. Abcam's AbpromiseSM

Quality is important to Abcam. If PDI antibody [RL90] - ER Marker (ab2792) does not perform as described on this datasheet, notify us within 90 days of delivery using this form, so that we can help you or offer you a replacement or a refund. See www.abcam.com/abpromise for more details.

Cellular localisation Endoplasmic reticulum lumen. Research areas Tags & Cell Markers >> Subcellular Markers >> Organelles >> ER Relevance The three dimensional structure of many extracellular proteins is stabilized by the formation of disulphide bonds. Studies suggest that a microsomal enzyme known as Protein Disulphide Isomerase (PDI) is involved in disulphide-bond formation and isomerization, as well as the reduction of disulphide bonds in proteins. PDI, which catalyses disulphide interchange between thiols and protein dilsulphides, has also been referred to as thiol:protein-disulphide oxidoreductase and as glutathione:insulin transhydrogenase because of its role in reduction of disulphide bonds. The highly conserved sequence Lys-Asp-Glu-Leu (KDEL) is present at the carboxy-terminus of PDI and other soluble endoplasmic reticulum (ER) resident proteins including the 78 and 94 kDa glucose regulated proteins (GRP78 and GRP94 respectively). The presence of carboxy-terminal KDEL appears to be necessary for ER retention and appears to be sufficient to reduce the secretion of proteins from the ER. This retention is reported to be mediated by a KDEL receptor. Alternative names for PDI antibody [RL90] - ER Marker (ab2792) Database links The links below go to external sites and will open in a new browser windowEntrez Gene 5034 (Human) 64714 (Human) 29943 (Human) 14827 (Mouse) 18453 (Mouse) 18600 (Mouse) 25506 (Rat)

Omim 176790 (Human)

SwissProt P07237 (Human) P09103 (Mouse) Q08642 (Mouse) P20717 (Rat) P04785 (Rat)

Unigene 410578 (Human) 16660 (Mouse) 4234 (Rat)

Raised in Mouse Clonality Monoclonal Clone number RL90 Isotype IgG2a Purity Ascites Storage buffer PBS with 0.05% sodium azide

Material safety datasheet (MSDS) for this product: Sodium Azide MSDS

Form Liquid Storage instructions Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.