Name: Anti-PKR antibody [YE350]
Brand: RabMAb
SKU: ab32052
Rating: 5 (1 reviews)

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Western blot - Anti-PKR antibody [YE350] (ab32052)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [YE350] to PKR
Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
Knockout validated
Reacts with: Human

Carrier Free

製品名

Anti-PKR antibody [YE350]
PKR 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [YE350] to PKR
由来種

Rabbit
特異性

This antibody recognises Protein kinase R (PKR). It does not cross-react with other GCN2 family members.
アプリケーション

適用あり: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details
種交差性

交差種: Human
免疫原

Synthetic peptide within Human PKR aa 50-150. The exact sequence is proprietary.
Database link: P19525
ポジティブ･コントロール
- WB: Jurkat, A549, K562, HAP1, HepG2, and HeLa cell lysates. IP: Jurkat cell lysat; IHC: Human cerebrum tissue; ICC/IF: MCF7 cells; Flow Cyt (intra): HeLa cells.
特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
バッファー

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

YE350
アイソタイプ

IgG
研究分野

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
KO cell lysates
Recombinant Protein
- Recombinant human PKR protein (ab71666)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab32052の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
Flow Cyt (Intra)		1/20 - 1/50.
WB	(1)	1/1000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
IP		1/20 - 1/100.
ICC/IF		1/50. For unpurified use at 1/100 - 1/500.

特記事項
Flow Cyt (Intra) 1/20 - 1/50.
WB 1/1000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
IP 1/20 - 1/100.
ICC/IF 1/50. For unpurified use at 1/100 - 1/500.

機能

Following activation by double-stranded RNA in the presence of ATP, the kinase becomes autophosphorylated and can catalyze the phosphorylation of the translation initiation factor EIF2S1, which leads to an inhibition of the initiation of protein synthesis. Double-stranded RNA is generated during the course of a viral infection.
配列類似性

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. GCN2 subfamily.
Contains 2 DRBM (double-stranded RNA-binding) domains.
Contains 1 protein kinase domain.
翻訳後修飾

Autophosphorylated on several Ser and Thr residues. Autophosphorylation of Thr-451 is dependent on Thr-446 and is stimulated by dsRNA binding and dimerization. Autophosphorylation apparently leads to the activation of the kinase.
Target information above from: UniProt accession P19525 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 5610 Human
- Omim: 176871 Human
- SwissProt: P19525 Human
- Unigene: 131431 Human
別名
- Double stranded RNA activated protein kinase; antibody
- E2AK2_HUMAN antibody
- eIF-2A protein kinase 2 antibody
- EIF2AK1 antibody
- EIF2AK2 antibody
- Eukaryotic translation initiation factor 2 alpha kinase 2 antibody
- Eukaryotic translation initiation factor 2-alpha kinase 2 antibody
- HGNC:9437 antibody
- Interferon induced double stranded RNA activated protein kinase antibody
- Interferon inducible elF2 alpha kinase antibody
- Interferon inducible RNA dependent protein kinase antibody
- Interferon-induced, double-stranded RNA-activated protein kinase antibody
- Interferon-inducible RNA-dependent protein kinase antibody
- MGC126524 antibody
- P1/eIF-2A protein kinase antibody
- P1/eIF2A protein kinase antibody
- p68 kinase antibody
- PKR antibody
- PPP1R83 antibody
- PRKR antibody
- Protein kinase interferon inducible double stranded RNA dependent antibody
- Protein kinase RNA activated antibody
- Protein kinase RNA-activated antibody
- Protein phosphatase 1 regulatory subunit 83 antibody
- Serine/threonine protein kinase TIK antibody
- Tyrosine protein kinase EIF2AK2 antibody
see all

Western blot - Anti-PKR antibody [YE350] (ab32052)

All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 5 : K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Lanes 1-5: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261824 (knockout cell lysate ab256899) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [YE350] (ab32052)

Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PKR with Purified ab32052 at 1:50 dilution (5.04 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [YE350] (ab32052)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling PKR with Purified ab32052 at 1:100 dilution (2.52 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Western blot - Anti-PKR antibody [YE350] (ab32052)

Anti-PKR antibody [YE350] (ab32052) at 1/5000 dilution (Purified) + Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Flow Cytometry (Intracellular) - Anti-PKR antibody [YE350] (ab32052)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKR with Purified ab32052 at 1/30 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunoprecipitation - Anti-PKR antibody [YE350] (ab32052)

Purified ab32052 at 1/20 dilution (1µg) immunoprecipitating PKR in Jurkat whole cell lysate.
Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg
Lane 2 (+): ab32052 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32052 in Jurkat whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 68 kDa
Possible degradation bands are observed between 20-30kDa.
Western blot - Anti-PKR antibody [YE350] (ab32052)

All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : EIF2AK2 knockout A549 cell lysate
Lane 3 : K-562 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267000 (knockout cell lysate ab256901) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-PKR antibody [YE350] (ab32052)

All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : EIF2AK2 knockout A549 cell lysate
Lane 3 : K-562 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266999 (knockout cell lysate ab256900) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-PKR antibody [YE350] (ab32052)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: EIF2AK2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: K652 whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab32052 was shown to specifically react with EIF2AK2 when EIF2AK2 knockout samples were used. Wild-type and EIF2AK2 knockout samples were subjected to SDS-PAGE. Ab32052 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

ab32052 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab32052 は 41 報の論文で使用されています。

Peng Z et al. PKR deficiency delays vascular aging via inhibiting GSDMD-mediated endothelial cell hyperactivation. iScience 26:105909 (2023). PubMed: 36691613
Chen S & Harris M NS5A domain I antagonises PKR to facilitate the assembly of infectious hepatitis C virus particles. PLoS Pathog 19:e1010812 (2023). PubMed: 36795772
Wang F et al. Novel lncRNA AL033381.2 Promotes Hepatocellular Carcinoma Progression by Upregulating PRKRA Expression. Oxid Med Cell Longev 2022:1125932 (2022). PubMed: 35035655
Song H et al. circFAM120B functions as a tumor suppressor in esophageal squamous cell carcinoma via the miR-661/PPM1L axis and the PKR/p38 MAPK/EMT pathway. Cell Death Dis 13:361 (2022). PubMed: 35436983
Werner A et al. Widespread formation of double-stranded RNAs in testis. Genome Res 31:1174-86 (2021). PubMed: 34158368

View all Publications for this product

レビューと Q&A

Show All レビュー Q&A

レビューを送る質問を送る

1-7 of 7 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa cell line)

Loading amount

20 µg

Specification

HeLa cell line

Gel Running Conditions

Reduced Denaturing (12% gel)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Mirjam Eckert

Verified customer

投稿 Mar 26 2009

Question

Could we get a detailed protocol from you, please. We have some general questions, the answers to which will enable us to investigate this matter as quickly as possible.
1. Order details:

Batch number:

Abcam product code: ab 32052

Antibody storage conditions (temperature/reconstitution etc): -20- deg C
2. Please describe the problem (high background, wrong band size, more bands, no band etc).
Two major bands were detected by the Ab; one with expected size, but the other one is much smaller.
3. On what material are you testing the antibody in WB?
· Species:
· What’s cell line or tissue: 293T
· Cell extract or Nuclear extract: Cell extract
· Purified protein or Recombinant protein:

3. The lysate

How much protein was loaded: ˜1 *10^5 cells/lane

What lysis buffer was used: directly lysed with SDS sample buffer

What protease inhibitors were used: no

What loading buffer was used: SDS sample buffer

Phosphatase inhibitors

Did you heat the samples: temperature and time: boil for 5 min

4. Electrophoresis/Gel conditions/ Transfer conditions

Reducing or non reducing gel:

Reducing agent:

Gel percentage : 12%

Transfer conditions: (Type of membrane, Protein transfer verified): NC membrane and wet transfer in 200mA for 90min

5. Blocking conditions

Buffer: PBS- 0.1% tween

Blocking agent: milk, BSA, serum, what percentage: 10% skim milk

Incubation time: 1 hour

Incubation temperature: room temperature

6. Primary Antibody

Species:

Reacts against:
· At what dilution(s) have you tested this antibody: 1:5000, 1:1000
· What dilution buffer was used: diluted in PBS-tween (5% skim milk)
· Incubation time: for overnight
· Incubation temperature: 4-deg C
· What washing steps were done: PBS-tween (5% skim milk) for more than 5 times.

7. Secondary Antibody

Species: Rabbit

Reacts against:

At what dilution(s) have you tested this antibody: 1:10000

Incubation time: 1hr

Wash steps: PBST 10min X2

Fluorochrome or enzyme conjugate: ECL

Do you know whether the problems you are experiencing come from the secondary?

8. Detection method
ECl, ECl+, other detection method: ECl

9. Did you apply positive and negative controls along with the samples? Please specify. PKR is endogenous protein. We don’t have an appropriate negative control for the expression of PKR.
10. Optimization attempts
· How many times have you tried the Western? Three times
· Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control):
· Do you obtain the same results every time e.g. are background bands always in the same place? yes
· What steps have you altered? Adjust the Ab concentration.

Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

Abcam community

Verified customer

Asked on Oct 09 2012

Answer

Thank you for your enquiry regardingab32052 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

Could you please confirm if you purchased this product directly from Abcam or via a distributor? It would be very useful for me to know the batch number and the date of the purchase.

It would be much appreciated if you could attach a new Western blot image which represents the MW (molecular weight markers)band and the descriptions of the different lanes. Without this information, it is impossible for us to interpret the WB image attached to this message.

I look forward to hearing from you soon.

Abcam Scientific Support

Answered on Oct 09 2012

Question

A few weeks ago you very kindly sent me a replacement PKR antibody as the new batch behaved very differently to previous batches (see email below). I have been away at a conference and so have not had a chance to confirm whether or not the new aliquot (same batch number) resolved the staining issues until now. Unfortunately this second aliquot from the same batch stains exactly the same as the last one. Therefore I believe that the issue is likely to be a batch one not an aliquot one. Could you please tell me when you might have a new batch of this antibody being produced? If this is likely to take a long period of time would you please consider sending us an alternative PKR antibody (suitable for IHC-P). I have looked through your selection and would be keen to try ab28943 if possible.

Abcam community

Verified customer

Asked on Mar 20 2012

Answer

Thank you for contacting Abcam. Unfortunately we are not aware of when the new batch will be available. I have arranged to send you the the replacement product (ab28943) that you have requested. 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 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.

Abcam Scientific Support

Answered on Mar 20 2012

Question

I would be grateful if you could send me a replacement from the same lot and I will screen this to determine if we were just unlucky with that particular viral. The previous order number for the last batch was: I look forward to receiving the replacement PKR antibody, Many thanks for your assistance. Best wishes

Abcam community

Verified customer

Asked on Feb 21 2012

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 1038340.

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.

Abcam Scientific Support

Answered on Feb 21 2012

Question

It is quite short but we use the DAKO antibody diluent and the Envision kit and have had no problems with background previously with other antibodies we use. In fact DAKO suggested we shouldn't need this step at all using the reagents as we do. As I mentioned earlier the two samples you are looking at we're both put through the same run so there is no difference between block times and the early batches (and ones before that) had no issues with background.new do not use an automated stainer. Ou samples are stained manually. I will email you the order number tomorrow once I return to work. I look forward to hearing from you, Best wishes,

Abcam community

Verified customer

Asked on Feb 20 2012

Answer

LOT:GR56038-1

I am sorry to confirm that we don't have any different lots in stock at the moment. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. Regrettably, I can suggest you have received a bad vial on this occasion.

Therefore, we would be pleased to provide a replacement from the same lot, which will still be covered by our guarantee. Alternatively, I fully understand your concerns and you prefer a credit note in this case, I will be pleased to arrange this for you.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

Abcam Scientific Support

Answered on Feb 20 2012

Question

Please find the details below you have requested. Tissue samples: Formalin fixed paraffin embedded human pancreas tissue. We have also screen in lymph node and other human tissues. HIER - 10mM citrate pH6 Normal goat serum block for 5 minutes PKR antibody - 1/350; initially we did titrations (1/100-1/1000) to optimise this. The antibody is incubated overnight at 4C. Detection system : Dako Envision HRP. Please let me know if you require further information.

Abcam community

Verified customer

Asked on Feb 20 2012

Answer

Thank you for providing these additional details.

I noted that your blocking time is low, a common period is between 30-60 minutes at room temperature. Do you use an automated staining system?

Would you be able to provide the Abcam order number or the PO number for this purchase?

Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Abcam Scientific Support

Answered on Feb 20 2012

Question

I am a regular user of your PKR antibody C#ab32052 and have been carrying out IHC and IF experiments in formalin-fixed paraffin embedded tissue with it for about 4-5 years now. We have published these results and are in the process of writing an additional manuscript using this antibody. At the end of last year I bought a new batch and am having problems with it. Below are images of the same islet in serial sections, the first was stained with an older batch and the second with the new batch. As you can see the staining pattern is completely different. These two were stained in parallel using the same buffers, concentrations and conditions. In particular the later batch appears to be labelling a subset of islet cells, a phenomenon that we have never seen in the 3-4 antibody batches we have purchased previously. There was no indication on the datasheet that the antibody concentration had been changed as far as I can determine. Please could you look into this and I would be grateful if you would consider sending us a different batch. I look forward to hearing from you soon, Best wishes Sarah Dr Sarah Richardson Post-doctoral Research Fellow Endocrine Pharmacology Institute of Biomedical and Clinical Science John Bull Building Peninsula Medical School Research Way Plymouth PL6 8BU Tel: 44 (0)1752 431038 FAX: 44 (0)1752 517846 E-mail: sarah.richardson@pms.ac.uk

Abcam community

Verified customer

Asked on Feb 15 2012

Answer

Thank you for contacting us.

I am sorry to hear of the difficulties that you have been experiencing using this product. Would you mind sending me the details of your protocol including the tissue type you are working with and your sample preparations? Could you also include any antigen retrieval steps you use, blocking solution and incubation information and the antibody concentrations that you have tried.

The images that you have provode are very different.It seems unlikely that your protocol is the issue here but these details will not only help me to better understand your difficulties, they are also used for internal testing of the products if deemed necessary. If we are unable to solve the problems you are experiencing we will replace or refund the antibody according to our Abpromise guarantee.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Feb 15 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

特異性

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

配列類似性

翻訳後修飾

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

参考文献 (41)

レビューと Q&A

Western blot abreview for Anti-PKR antibody [YE350]