Overview

  • Product name
    Anti-PRDM1/Blimp1 antibody - ChIP Grade
    See all PRDM1/Blimp1 primary antibodies
  • Description
    Goat polyclonal to PRDM1/Blimp1 - ChIP Grade
  • Host species
    Goat
  • Tested applications
    Suitable for: Flow Cyt, ChIP, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow, Dog, Pig
  • Immunogen

    Synthetic peptide: KVKQETVEPMDP, corresponding to C terminal amino acids 778-789 of Human PRDM1/Blimp1.

  • Positive control
    • Human Lymph Node lysate
  • General notes
    GenBank Accession Number - NP_001189

Properties

Applications

Our Abpromise guarantee covers the use of ab13700 in the following tested applications.

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

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration.

ab37373 - Goat polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

ChIP Use at an assay dependent concentration. PubMed: 17264218
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 91 kDa).

Target

  • Function
    Transcriptional repressor that binds specifically to the PRDI element in the promoter of the beta-interferon gene (PubMed:1851123). Drives the maturation of B-lymphocytes into Ig secreting cells (PubMed:12626569).
  • Sequence similarities
    Belongs to the class V-like SAM-binding methyltransferase superfamily.
    Contains 4 C2H2-type zinc fingers.
    Contains 1 SET domain.
  • Post-translational
    modifications
    Sumoylation at Lys-816 by PIAS1 augments transcriptional repressor activity, and is critical for plasma cell differentiation.
    Ubiquitinated by the SCF(FBXO11) complex, leading to its degradation by the proteasome.
  • Cellular localization
    Nucleus. Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • B Lymphocyte Induced Maturation Protein 1 antibody
    • Beta interferon gene positive regulatory domain I binding factor antibody
    • Beta-interferon gene positive regulatory domain I-binding factor antibody
    • BLIMP-1 antibody
    • BLIMP1 antibody
    • Positive Regulatory Domain I Binding Factor 1 antibody
    • Positive regulatory domain I-binding factor 1 antibody
    • PR Domain Containing 1 antibody
    • PR domain containing 1 with ZNF domain antibody
    • PR domain containing 1 with ZNF domain isoform 2 antibody
    • PR domain containing protein 1 antibody
    • PR domain zinc finger protein 1 antibody
    • PR domain-containing protein 1 antibody
    • PRDI BF1 antibody
    • PRDI binding factor 1 antibody
    • PRDI-BF1 antibody
    • PRDI-binding factor 1 antibody
    • PRDM 1 antibody
    • Prdm1 antibody
    • PRDM1_HUMAN antibody
    see all

Images

  • Ab13700 staining (1µg/ml) of human Lymph Node lysate (RIPA buffer, 35µg total protein per lane).  Primary incubated for 1 hour.  Detected by western blot using chemiluminescence.

  • Anti-PRDM1/Blimp1 antibody - ChIP Grade (ab13700) at 1 µg/ml (for 1 hour) + Human brain (cerebellum) lysate in RIPA buffer at 35 µg

    Predicted band size: 91 kDa

  • Human RPMI8226 cells stained with ab13700 at 5 µg/ml, incubated for 30 minutes at 4°C.  In parallel, cells were stained with goat IgG at an equivalent concentration to that of PRDM1. The secondary antibody was a FITC-conjugated rabbit anti-goat IgG F(ab)2.  PRDMI expression was analysed by flow cytometry.

    See Abreview

References

This product has been referenced in:
  • Zhao H  et al. GPR39 marks specific cells within the sebaceous gland and contributes to skin wound healing. Sci Rep 5:7913 (2015). IF . Read more (PubMed: 25604641) »
  • Beermann ML  et al. Prdm1 (Blimp-1) and the expression of fast and slow myosin heavy chain isoforms during avian myogenesis in vitro. PLoS One 5:e9951 (2010). Read more (PubMed: 20376350) »
See all 5 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Chicken Cell lysate - whole cell (293FT transfected with expression vector, DT40)
Gel Running Conditions
Reduced Denaturing (7.5%)
Loading amount
10 µg
Specification
293FT transfected with expression vector, DT40
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Feb 26 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, embryonic day 15.5)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, embryonic day 15.5
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde

Mr. Bryan Niedenberger

Verified customer

Submitted Apr 28 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Sample
Pig Cell (Gonad day 25 fetus)
Specification
Gonad day 25 fetus
Permeabilization
Yes - citrate antigen retrival
Fixative
Paraformaldehyde

Dr. Ramiro Alberio

Verified customer

Submitted Jul 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (RPMI8226 cell line)
Specification
RPMI8226 cell line
Preparation
Cell harvesting/tissue preparation method: The cells were harvested and stained with 7AAD for 20 minutes on ice in the dark before the cells were fixed and permeabilised using the BD Cytofix/Cytoperm kit. Intracellular staining for PRDM1 was performed in Perm/wash in the presence of 10% human AB serum for 30 minutes on ice in the dark. In parallel, cells were stained with goat IgG at an equivalent concentration to that of PRDM1. Following three washes in Perm/wash the cells were stained with FITC-conjugated rabbit anti-goat IgG for 30 min on ice in the dark. Following three washes PRDMI expression was analysed by flow cytometry.
Sample buffer: BD Cytofix/Cytoperm
Fixation
Formaldehyde
Permeabilization
Yes - BD Cytofix/Cytoperm Kit
Gating Strategy
All live RPMI8226 cells were included in the analysis. Dead cells were excluded in FL-3 using 7AAD.

Dr. Angelica Cazaly

Verified customer

Submitted Dec 03 2007

Answer

Thank you for your enquiry. We do not have any data that the antibody shows the 2 isoforms on western blots of myeloma cell lysates. We only have the data on the lymphoma specimen. As long as the amino acids 778-789 (KVKQETVEPMDP) are present in both isoforms and the proteins can be separated sufficiently on an SDS-PAGE gel, then I do not see why this antibody should not be able to detect both isoforms. Of course, as I mentioned, we have not formally tested this. I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

Read More

Question
Answer

I have received a nice reply from the supplier of the ICC image. He states "We simply used a very standard protocol. Fixed for 20 mins at 4C in 4%PFA, blocked in 10% donkey serum for about 60 mins at room temp, then added primary overnight at 4C, then secondary for 4 hours at 4C". I hope this information helps. Please do not hesitate to contact me should you require further assistance.

Read More

Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with ab13700. The ICC image on the datasheet was submitted by a customer who wishes to remain anonymous. I have contacted him directly requesting further information if there was anything out of the ordinary done to get this antibody working in ICC. I will contact you in due course should any details emerge. With regards to a positive control, following some literature searching I came across: Chang DH, Angelin-Duclos C, Calame K. BLIMP-1: trigger for differentiation of myeloid lineage. Nat Immunol. 2000 Aug;1(2):169-76. In the paper they treated promyeloid leukemia cell lines U937 and HL60. By inducting them to differentiate using PMA they directly upregulated BLIMP1, detectable using ICC and the ribonuclease protection assay. Therefore I think that if you could get hold of one of these cell lines and perhaps monitor their BLIMP1 mRNA levels to confirm upregulation they would serve as excellent positive controls. Should this not be of assistance I would encourage you to submit a technical questionnaire by clicking on the link below. This will better give us an appreciation of the optimisation steps that you have undertaken. https://www.abcam.com/index.html?section=ihc&pageconfig=technical&intAbID=13700&mode=questionaire

Read More

Answer

Thank you for your reply and for sending us further information about your customer's Western blot protocol. Looking through the original completed Questionnaire and the forwarded extra-information you have just sent to us, I would like to make the following suggestions: As the datasheet of this antibody indicates, ab13700 has been tested and characterised in human lymph node lysate. 35µg total protein per lane of this type of lysate was used (prepared in RIPA buffer) and the antibody at a concentration of 1µg/ml was incubated with the membrane. Unfortunately, we do not have information about the cell line (RPMI 8226) your customer is using. Therefore we would strongly suggest using human lymph node lysate as a well characterised positive control. For Western blot application, it is extremely important to determine the protein concentration of the samples. The customer should load 20-30 total protein per lane. The lysis buffer is also important, and we would recommend your customer using RIPA buffer: Preparation of RIPA cell lysates for Western Blots: 1. Wash cell pellets once with ice-cold PBS. 2. Add 1 ml of RIPA buffer to 10x8 cells, incubate on ice for 20 min, vortex 2 to 3 times. 3. Centrifuge for 5 min at 4oC at maximum speed in a microfuge tube. 4. Transfer supernatant into clean tube. Measure protein concentration with a protein assay. 5. Adjust concentration to 5 mg/ml with RIPA lysis buffer. 6. Add equal volume of 2 x SDS sample buffer into cell lysate, boil for 5 min. 7. Store at -20oC for daily use or -80oC for long term. Avoid repeated freeze thaw cycles. RIPA Base Ingredients Tris-HCl: 50 mM, pH 7.4 NP-40: 1% Na-deoxycholate: 0.25% NaCl: 150 mM EDTA: 1 mM PMSF: 1 mM Aprotinin, leupeptin, pepstatin: 1 microgram/ml each RIPA Protease Inhibitors Phenylmethylsulfonyl fluoride (PMSF) (200 mM stock solution in isopropanol; store at room temperature) EDTA (calcium chelator; 100 mM stock solution in H2O, pH 7.4) Leupeptin (store frozen in aliquots, 1 mg/ml in H2O) Aprotinin (store frozen in aliquots, 1 mg/ml in H2O) Pepstatin (store frozen in aliquots, 1 mg/ml in methanol) We hope this information will be useful.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up