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    grp78-bip-antibody-ab32618.pdf

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Anti-GRP78 BiP antibody (ab32618)

  • Datasheet
  • SDS
Submit a review Q&A (9)References (43)

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Western blot - Anti-GRP78 BiP antibody (ab32618)
  • Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab32618)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab32618)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab32618)

Key features and details

  • Rabbit polyclonal to GRP78 BiP
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Human, Chinese hamster
  • Isotype: IgG

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Overview

  • Product name

    Anti-GRP78 BiP antibody
    See all GRP78 BiP primary antibodies
  • Description

    Rabbit polyclonal to GRP78 BiP
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Human
    Chinese hamster
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human GRP78 BiP aa 1-100. The exact sequence is proprietary.
    Database link: P11021

  • Positive control

    • HeLa cells, breast carcinoma.
  • General notes

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.60
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • ER

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • Recombinant Human GRP78 BiP protein (ab79139)
  • Recombinant Protein

    • Recombinant human GRP78 BiP protein (Active) (ab78432)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32618 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Mouse
Human
Chinese hamster
All applications
Rat
Chicken
Hamster
Xenopus laevis
Application Abreviews Notes
WB
Use at an assay dependent concentration. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa).
IHC-P
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
1/100.
Notes
WB
Use at an assay dependent concentration. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa).
IHC-P
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
1/100.

Target

  • Function

    Probably plays a role in facilitating the assembly of multimeric protein complexes inside the endoplasmic reticulum. Involved in the correct folding of proteins and degradation of misfolded proteins via its interaction with DNAJC10, probably to facilitate the release of DNAJC10 from its substrate.
  • Involvement in disease

    Autoantigen in rheumatoid arthritis.
  • Sequence similarities

    Belongs to the heat shock protein 70 family.
  • Cellular localization

    Endoplasmic reticulum lumen. Melanosome. Cytoplasm. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Target information above from: UniProt accession P11021 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 396487 Chicken
    • Entrez Gene: 100689305 Chinese hamster
    • Entrez Gene: 3309 Human
    • Entrez Gene: 14828 Mouse
    • Entrez Gene: 25617 Rat
    • Entrez Gene: 397850 Xenopus laevis
    • Omim: 138120 Human
    • SwissProt: Q90593 Chicken
    • SwissProt: P07823 Hamster
    • SwissProt: P11021 Human
    • SwissProt: P20029 Mouse
    • SwissProt: P06761 Rat
    • SwissProt: Q91883 Xenopus laevis
    • Unigene: 743241 Human
    • Unigene: 330160 Mouse
    • Unigene: 470180 Mouse
    • Unigene: 474909 Mouse
    • Unigene: 11088 Rat
    • Unigene: 3204 Xenopus laevis
    see all
  • Alternative names

    • 78 kDa glucose regulated protein antibody
    • 78 kDa glucose-regulated protein antibody
    • AL022860 antibody
    • AU019543 antibody
    • BIP antibody
    • D2Wsu141e antibody
    • D2Wsu17e antibody
    • Endoplasmic reticulum lumenal Ca(2+)-binding protein grp78 antibody
    • Endoplasmic reticulum lumenal Ca2+ binding protein grp78 antibody
    • Epididymis secretory sperm binding protein Li 89n antibody
    • FLJ26106 antibody
    • Glucose Regulated Protein 78 antibody
    • Glucose Regulated Protein 78kDa antibody
    • GRP 78 antibody
    • GRP-78 antibody
    • GRP78 antibody
    • GRP78_HUMAN antibody
    • Heat shock 70 kDa protein 5 antibody
    • Heat Shock 70kDa Protein 5 antibody
    • Heat shock protein family A (Hsp70) member 5 antibody
    • HEL S 89n antibody
    • Hsce70 antibody
    • HSPA 5 antibody
    • HSPA5 antibody
    • Immunoglobulin Heavy Chain Binding Protein antibody
    • Immunoglobulin heavy chain-binding protein antibody
    • mBiP antibody
    • MIF2 antibody
    • Sez7 antibody
    see all

Images

  • Western blot - Anti-GRP78 BiP antibody (ab32618)
    Western blot - Anti-GRP78 BiP antibody (ab32618)
    All lanes : Anti-GRP78 BiP antibody (ab32618) at 1 µg/ml

    Lane 1 : Liver (Mouse) Tissue Lysate
    Lane 2 : CHO-K1 cell lysate Whole Cell Lysate
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 78 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    The band observed at 75 kDa could potentially be a cleaved form of GRP78 BiP due to the presence of a 18 amino acid signal peptide.
  • Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab32618)
    Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab32618)
    ICC/IF image of ab32618 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32618, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab32618)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab32618)
    This image shows human breast carcinoma stained with ab32618 diluted 1/100.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab32618)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab32618)
    Ab32618 staining Human normal liver parenchyma. Staining is localised to endoplasmic reticulum compartment.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be requi

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (43)

Publishing research using ab32618? Please let us know so that we can cite the reference in this datasheet.

ab32618 has been referenced in 43 publications.

  • Bandla S  et al. ATPase activity of human binding immunoglobulin protein (BiP) variants is enhanced by signal sequence and physiological concentrations of Mn2. FEBS Open Bio 9:1355-1369 (2019). PubMed: 31033254
  • Guan G  et al. Mechanism of interactions between endoplasmic reticulum stress and autophagy in hypoxia/reoxygenation-induced injury of H9c2 cardiomyocytes. Mol Med Rep 20:350-358 (2019). PubMed: 31115545
  • Lei L  et al. Role of endoplasmic reticulum stress in lipopolysaccharide-inhibited mouse granulosa cell estradiol production. J Reprod Dev 65:459-465 (2019). PubMed: 31406023
  • Wang K  et al. Role of GRP78 inhibiting artesunate-induced ferroptosis in KRAS mutant pancreatic cancer cells. Drug Des Devel Ther 13:2135-2144 (2019). PubMed: 31456633
  • Chen Z  et al. Curcumin protects against palmitic acid-induced apoptosis via the inhibition of endoplasmic reticulum stress in testicular Leydig cells. Reprod Biol Endocrinol 17:71 (2019). PubMed: 31472681
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Question

tatsächlich hatten wir Probleme bei der Anwendung des Bestellten Antikörpers
(ab32618) in unserem Labor.
Während es bei der Anwendung auf Hefeproteine nur zu sehr schwachen Signalen
kam konnte bei der Anwendung auf Algenproteine in keinem Fall ein Signal
detektiert werden (Western Blot).
Letzteres hat jedoch mit früheren Chargen von ab32618 problemlos
funktioniert. Allerdings weicht die batch number von der von Ihnen
angegebenen Nummer ab (GR19355-2). Es ist für uns daher nicht klar, ob die
von uns beobachtete Problematik mit den von Ihnen beschriebenen
Qualitätsproblemen von GR67285 zusammenhängt.

Read More

Abcam community

Verified customer

Asked on Oct 19 2012

Answer

Vielen Dank fuer Ihre email.
Es tut mir leid zu hoeren, dass Sie Problem mit diesem Antikoerper haben.

Die Information bezueglich des defekten Lots GR67285 bezieht sich tatsaechlich nur auf dieses Lot.

Da wir keine Daten fuer Hefe- und Algenproben auf unserem Datenblatt haben, koennen wir den Antikoerper fuer diese Spezien nicht garantieren. Ich verstehe, dass fruehere Lots fuer Sie funktioniert haben.

Ich wuerde daher vorschlagen, dass Sie am unserem Abtrail program teilnehmen, da Hefe und Algen fuer uns ungetestete Spezien sind. Sie koennen jeweils 1 Abreview fuer jede Spezie mit den positiven Daten einreichen, und dann einen Rabatt fuer den Preis des Antikorpers erhalten, der dann von einer neuer Bestellung abgezogen werden kann. Sobald wir die positiven Daten publiziert haben auf unserem Datenblatt, werden die Spezien Hefe und Alge als getestete Spezien hinzugefuegt.

Hier sind die genauen Informationen bezueglich des Abtrial Programs:

For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein/kit on an untested species/application. You would purchase the to-be-tested product at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount in the amount of the price you paid for the to-be-tested product. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/abtrial.


Bitte sagen Sie mir doch Bescheid, ob ich Ihnen einen Rabattkode (discount code) zustellen soll.

Ich hoffe, dass Ihnen das weiterhelfen wird.

Read More

Abcam Scientific Support

Answered on Oct 19 2012

Question

feedback in the follow up

Read More

Abcam community

Verified customer

Asked on Sep 14 2012

Answer

Thank you very much for having provided your feedback.

I just have tried to call you, however I have been told you are in vacation.

I am very sorry that it took so long for solving the problem in this case. When I sent the first replacement vial, I did not know it was due to the batch - as the problem could have just come from the shipping. This is why I did not request the laboratory whether there is soon a new batch available or not. I have now requested the laboratory to investigate this batch again and retest it.

I believe we have replaced all the vials to you which were from the faulty batch. Please do let me however know if this is not the case.

In order to apologize for all the inconvenience, we are pleased to offer you an additional free of charge vial of this antibody. Please do let us know whether you would like to accept this.

Thank you for your collaboration. We look forward to hear back from you.

Read More

Abcam Scientific Support

Answered on Sep 14 2012

Question

We have tried the new batch of GRP 78 and it worked very well. It is a pitty that it took this long to solve the problem, time and money. We have the 3 vials that did not work, we can send themback to you. Also we think that it would be fair for ABCAM to ship another vial of the same batch free of charge.
Best Regards

Read More

Abcam community

Verified customer

Asked on Sep 04 2012

Answer

Thank you for your email.

I am happy to hear that the new batch provides again the expected results. Also, I am very sorry for all the troubles encountered. We will replace all the vials of the old batch for you with a vial of the new batch. Can you please provide me with all the order numbers of the ab32618 you placed, with the date? I will then check how many vials we will be able to send you/reimbourse you.

Thank you very much for your cooperation. I look forward to your reply.

Read More

Abcam Scientific Support

Answered on Sep 04 2012

Question

Could you please send us now the new batch ofGRP78 antibody, we are back from holidays and would like to give it a try.

Best Regards

Read More

Abcam community

Verified customer

Asked on Aug 28 2012

Answer

Thank you for your email.


I have issued a free of charge replacement of a vial of the new batch of ab32618 with the order number x. To check the status of the order please contact our Customer Service team and reference this number.

I hope that this batch will provide finally good results again. I apologize for all the inconvenience. Please keep us updated on the results obtained with this vial.

I wish you a great day.

Read More

Abcam Scientific Support

Answered on Aug 28 2012

Question

I regret to inform you that once again the antibody did not work (the results are attached).We would like to know when will, yoube having a new batch. xx is on holidays, when he comes back he probably will be interested in an alternative solution, therefore he can discuss that with you. Also we would like to ask for a credit note on the antibody.

Best Regards

Read More

Abcam community

Verified customer

Asked on Aug 08 2012

Answer

Thank you for your email. I am very sorry to hear that the replacement vial has not provided the expected results. We do not know the reason of this problem yet, but we will investigate this. Please do also keep us updated if you have new information in regards to the antibodies or the samples used.

We have now received a new lot in stock of this antibody in the UK. Please let me know if you would like to wait for the return of xx to know whether you would like a replacement with the new lot or if you prefer the credit note.

Thank you for your cooperation. I look forward to your reply.

Read More

Abcam Scientific Support

Answered on Aug 08 2012

Question


We also found these results very awkward since this particular antibody always worked just fine until now. I would be grateful if you could send us one vial of GRP78 and replace the other vial with a credit note/refund.

Thank you for your cooperation and understanding,

Read More

Abcam community

Verified customer

Asked on Jul 27 2012

Answer

Thank you for your reply. As requested, I have requested a credit note and arranged the sending of the replacement vial. Please find here the credit note details. The others will follow.

Your credit note ID is xx


I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Read More

Abcam Scientific Support

Answered on Jul 27 2012

Question

We have been using a well-known “Protease Inhibitor Mix” from GE Healthcare, as mentioned in the questionnaire I sent you. This cocktail targets Calpain II, Cathepsin B, Elastase, leukocyte, Elastase, pancreatic, Papain, Plasmin, Thermolysin, Trypsin. Thus, the protein content of sample lysates, including GRP78, should not be degraded.

We have tested both GRP78 vials, more than three times each, without success.

Thank you.

Best regards,

Read More

Abcam community

Verified customer

Asked on Jul 25 2012

Answer

Thank you for your answer.

I am very sorry that I missed the line about the protease inhibitors. I have myself never worked with stomach and I do not know how the lysate is susceptible to degradation. We will send you replacement vials, however I would nevertheless appreciate if you can provide a positive control for the presence of GRP78 BiP in the samples tested with these new vials. A positive control can for example be Hela cell lysate.

I hope you understand that the product quality is very important and that we want therefore to exclude any other problems. We had not received any complaints in regards to this antibody in the last few years neither. I can however not exclude that your vials have been damaged during the transport or storage for example.

Please let me know how you would like to proceed. We can offer either free replacement vials or a credit note/refund. We do have currently no other lots of this antibody in stock, however as I said, we had not receive any other negative feedback on this lot neither.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Abcam Scientific Support

Answered on Jul 25 2012

Question

Dear Sirs,

Following previous contacts, please see file in attachment.

Best regards,

Read More

Abcam community

Verified customer

Asked on Jul 24 2012

Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that the new vials arenot providing satisfactory results.

Having reviewed this case, I would also appreciate if you can confirm some further details:

1.) I noticed that you have not used protease inhibitors in the samples. Can you confirm that the samples do contain still the GRP78 BIP protein? It could be that meanwhile the protein has been degraded. Especially when working with stomach, I would recommend to test freshly prepared samples, containing protease inhibitor cocktail

2.) Have you tested both vials you have ordered or have you just tested one of the two vials?
.
As already confirmed, if you can confirm that the protein is still present in the samples at the time of test, we will replace the vials for you according to our Abpromise.

I hope this information is helpful, and I thank you for your cooperation. I look forward to hear back from you.

Read More

Abcam Scientific Support

Answered on Jul 24 2012

Question


Hi I am writing to you regarding a purchase we made of GRP78cat# 63618lot#xxx.

We have been using GRP78 for immunoblots for quite a while, the one that we purchased in Febuary worked very well as seen on the picture atached (B). When we ordered more GRP78 in June, we saw that it was the same batch, so we decided to order another one for the work we had lined up. When we usedboth GRP78 ordered in June we found that there was no signal at all, as seen in the picture attached (A). We would like to know if you have another batch that we can try or solution to this problem.

Thank You

Read More

Abcam community

Verified customer

Asked on Jul 19 2012

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with the new vials ofthe ab32618 (I guess it is ab32618 you ordered, as ab63618 does not correspond to the description). We have received no other negative feedback so far and we are not aware that anything might has gone wrong during storage. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

We do unfortunately not have any other batch in stock for the moment. I would however appreciate if we caninvestigate this issue further and would appreciate if you could provide us some more information. Can you send us maybe more details on the protocol which has been used (with our questionnairehere attached - it just should take you 5 minutes to fill it out)? Can you please also confirm, that no other reactifs have been changed meanwhile, such as the secondary antibody for example? Thank you for having sent already the image with the result.

I would like to reassure you, that we do guarantee all our products to work as stated on the datasheet. Should the vials you have received been damaged, we would be happy to replace them for you.

Thank you very much for your cooperation. I look forward to receiving your reply.

Read More

Abcam Scientific Support

Answered on Jul 19 2012

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