Anti-Blooms Syndrome Protein Blm antibody (ab2179)

Product name

Anti-Blooms Syndrome Protein Blm antibody
See all Blooms Syndrome Protein Blm primary antibodies
Description

Rabbit polyclonal to Blooms Syndrome Protein Blm
Host species

Rabbit
Tested applications

Suitable for: WB, IP, ICCmore details
Species reactivity

Reacts with: Human
Predicted to work with: Cow, Dog, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan
Immunogen

Synthetic peptide (the amino acid sequence is considered to be commercially sensitive).

Form

Liquid
Storage instructions

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

Preservative: 0.1% Sodium azide
Concentration information loading...
Purity

Immunogen affinity purified
Clonality

Polyclonal
Isotype

IgG
Research areas

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

Our Abpromise guarantee covers the use of ab2179 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
WB		1/1000 - 1/10000. Detects a band of approximately 170 kDa (predicted molecular weight: 159 kDa).
IP		Use a concentration of 1 - 4 µg/ml.
ICC		Use a concentration of 5 µg/ml.

Function

Participates in DNA replication and repair. Exhibits a magnesium-dependent ATP-dependent DNA-helicase activity that unwinds single- and double-stranded DNA in a 3'-5' direction.
Involvement in disease

Defects in BLM are the cause of Bloom syndrome (BLM) [MIM:210900]. BLM is an autosomal recessive disorder characterized by proportionate pre- and postnatal growth deficiency, sun-sensitive telangiectatic hypo- and hyperpigmented skin, predisposition to malignancy, and chromosomal instability.
Sequence similarities

Belongs to the helicase family. RecQ subfamily.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain.
Contains 1 HRDC domain.
Post-translational
modifications

Phosphorylated in response to DNA damage. Phosphorylation requires the FANCA-FANCC-FANCE-FANCF-FANCG protein complex, as well as the presence of RMI1.
Cellular localization

Nucleus.
Target information above from: UniProt accession P54132 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 641 Human
- Omim: 604610 Human
- SwissProt: P54132 Human
- Unigene: 725208 Human
Alternative names
- Blm antibody
- BLM_HUMAN antibody
- Bloom syndrome antibody
- Bloom syndrome protein antibody
- Bloom syndrome RecQ helicase like antibody
- BS antibody
- DNA helicase antibody
- DNA helicase RecQ like type 2 antibody
- MGC126616 antibody
- MGC131618 antibody
- MGC131620 antibody
- RECQ 2 antibody
- RECQ like antibody
- RecQ like type 2 antibody
- RecQ protein like 3 antibody
- RecQ protein-like 3 antibody
- RecQ-like type 2 antibody
- RecQ2 antibody
- RECQL 2 antibody
- RECQL 3 antibody
- RECQL2 antibody
- RECQL3 antibody
- type 2 antibody
see all

Western blot - Anti-Blooms Syndrome Protein Blm antibody (ab2179)

All lanes : Anti-Blooms Syndrome Protein Blm antibody (ab2179)

Lane 1 : HeLa cells
Lane 2 : 293T cells
Lane 3 : Jurkat cells

Lysates/proteins at 50 µg per lane.

Predicted band size: 159 kDa

Detection:Chemiluminescence

Exposure time:30 seconds
Immunocytochemistry - Anti-Blooms Syndrome Protein Blm antibody (ab2179)This image was submitted as part of a review by Kirk McManus, University of British Columbia

HeLa cells cultured on coverslips were fixed in 4% paraformaldehyde then stained with ab2179 (green) at a concentration of 5µg/ml. The DNA stained with DAPI is shown in red (100x magnification).
Western blot - Anti-Blooms Syndrome Protein Blm antibody (ab2179)
Immunocytochemistry - Anti-Blooms Syndrome Protein Blm antibody (ab2179)This image was submitted as part of a review by Kirk McManus, University of British Columbia

HeLa cells were treated with 15µM Bleomycin for 2 hours to generate DNA double stranded breaks (DSBs). Paraformaldehyde-fixed cells were co-immunofluorescently labelled with anti-gamma H2AX (DNA DSB marker) and anti-BLM (ab2179). ab 2179 was used at a concentration of 5µg/ml. Colocalization at the sites of DNA DSBs is identified by the arrows. In the final panel DAPI is pseudo-coloured blue, while anti-gamma H2AX and anti-Blm are green and red respectively.

Western blot protocols
Immunoprecipitation protocols
Immunocytochemistry & immunofluorescence protocols

This product has been referenced in:

Gong Y et al. PHF11 promotes DSB resection, ATR signaling, and HR. Genes Dev 31:46-58 (2017). WB ; Mouse . Read more (PubMed: 28115467) »
Biehs R et al. DNA Double-Strand Break Resection Occurs during Non-homologous End Joining in G1 but Is Distinct from Resection during Homologous Recombination. Mol Cell 65:671-684.e5 (2017). Read more (PubMed: 28132842) »

See all 19 Publications for this product

Publishing research using ab2179? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 17 Abreviews or Q&A

Application

Immunoprecipitation

Sample

Human Cell lysate - whole cell (hek293)

Total protein in input

300 µg

Immuno-precipitation step

Other - protein G dynabeads, 10 microliters

Specification

hek293

Dr. Edyta Marcon

Verified customer

Submitted Apr 30 2015

Application

Western blot

Loading amount

50 µg

Gel Running Conditions

Non-reduced Denaturing (8%)

Sample

Hamster Cell lysate - whole cell (Chinese Hamster Ovary cells (TRex CHO ))

Specification

Chinese Hamster Ovary cells (TRex CHO )

Blocking step

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

Dr. Eleni Petsalaki

Verified customer

Submitted Feb 05 2014

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - whole cell (NTERA-2)

Loading amount

25 µg

Specification

NTERA-2

Gel Running Conditions

Reduced Denaturing (4-12%)

Abcam user community

Verified customer

Submitted Feb 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - whole cell (ntera2)

Loading amount

60000 cells

Specification

ntera2

Treatment

siRNA of Bloom

Gel Running Conditions

Reduced Denaturing

Blocking step

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

Mr. Faisal Alzahrani

Verified customer

Submitted Oct 18 2012

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (Colon carcinoma cells)

Specification

Colon carcinoma cells

Fixative

Methanol

Permeabilization

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C

Dr. Eleni Petsalaki

Verified customer

Submitted Oct 08 2012

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Loading amount

50 µg

Gel Running Conditions

Non-reduced Denaturing (8% gel)

Sample

Human Cell lysate - whole cell (Colon carcinoma)

Specification

Colon carcinoma

Blocking step

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

Dr. Eleni Petsalaki

Verified customer

Submitted Oct 08 2012

Question

DESCRIPTION OF THE PROBLEM Non-specific staining
SAMPLE Immortalized Human Bloom Syndrome cell line (6040 hTERT)
PRIMARY ANTIBODY ab2179 dilutions, 1:100, 1:200, and 1:400 incubated 1hr, wash: 3X 5 min in 0.2% tween-20 in pbs
DETECTION METHOD Fluorescent microscope
POSITIVE AND NEGATIVE CONTROLS USED Positive Control: BJ hTERT (WT) Negative Control: 6040 hTERT (BS line)
ANTIBODY STORAGE CONDITIONS 4C
FIXATION OF SAMPLE 4% Paraformaldehyde
PERMEABILIZATION STEP 1min in .2% Tween-20 in PBS
BLOCKING CONDITIONS Blocking Buffer: 1% BSA, 0.2% tween-20, 0.02% azide in PBS 30 min at 37C
SECONDARY ANTIBODY Invitrogen: Alexa Fluor 488 goat anti-rabbit IgG 1:400 dilution, 1hr at 37C wash: 3X 5 min in 0.2% tween-20 in pbs
HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1
HAVE YOU RUN A NO PRIMARY CONTROL? Yes
DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes
ADDITIONAL NOTES I've attached a picture of the IF I obtained on my BS cell line (negative control) Blue: dapi, Green: Blm ab2179. I've looked through the literature and haven't been able to find anyone using any of the BLM antibodies against a BS cell line. Has this or any of the BLM antibodies been tested on Bloom Syndrome cell lines?

Abcam community

Verified customer

Asked on Mar 13 2012

Answer

Thank you for contacting Abcam regarding ab2179.

I am sorry that you have been experiencing difficulties with this antibody in ICC. I have reviewed the protocol information you provided and would like to make some suggestions to improve your results with this antibody.

Regarding your question about your cells, at this time we have not tested nor received feedback from anyone who has used this antibody in a Bloom Syndrome cell line. As a positive control, I would recommend testing HeLa cells treated with 15uM Bleomycin for 2 hours as this produces a positive result with the antibody.

In terms of protocol modifications, I would recommend the following:

Since this is a nuclear protein, I would recommend triton X-100 to permeabilize - try 0.1% in PBS for 10 min. If you continue to use Tween-20 I would increase this to at least 10 min. Alternatively I would recommend fixation with acetone, which will both fix and permeabilize the cells.

The blocking step seems sufficient, but I would increase the incubation with the primary antibody to at least 2 hours, although ideally overnight at 4oC.

I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or if these suggestions do not improve your results with this antibody. The antibody is guaranteed under our Abpromise for WB and I would be happy to offer a replacement or credit if we determine the antibody is not working properly.

Abcam Scientific Support

Answered on Mar 13 2012

Application

Western blot

Sample

Mouse Cell lysate - whole cell (mouse testis)

Loading amount

100 µg

Specification

mouse testis

Treatment

Irradiation - 20 gy

Gel Running Conditions

Reduced Denaturing (8% Precast gel (Invitrogen))

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jan 12 2012

Application

Immunocytochemistry

Sample

Mouse Cell (mouse testis)

Specification

mouse testis

Fixative

Paraformaldehyde

Permeabilization

Blocking step

Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jan 12 2012

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunocytochemistry

Sample

Human Cell (HEK293-T)

Specification

HEK293-T

Fixative

Paraformaldehyde

Permeabilization

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Dec 05 2011

1-10 of 17 Abreviews or Q&A

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Anti-Blooms Syndrome Protein Blm antibody (ab2179)

Overview

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References

This product has been referenced in:

Customer reviews and Q&As

Immunoprecipitation abreview for Anti-Blooms Syndrome Protein Blm antibody

Western blot abreview for Anti-Blooms Syndrome Protein Blm antibody

Western blot abreview for Anti-Blooms Syndrome Protein Blm antibody

Western blot abreview for Anti-Blooms Syndrome Protein Blm antibody

Immunocytochemistry/ Immunofluorescence abreview for Anti-Blooms Syndrome Protein Blm antibody

Western blot abreview for Anti-Blooms Syndrome Protein Blm antibody

Western blot abreview for Anti-Blooms Syndrome Protein Blm antibody

Immunocytochemistry abreview for Anti-Blooms Syndrome Protein Blm antibody

Immunocytochemistry abreview for Anti-Blooms Syndrome Protein Blm antibody