SOCS3 antibody (ab16030)

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Anonymous Abreview submitted on 15 February 2009
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
SOCS3 antibody (ab16030)
Sample	Human Tissue sections (human term placenta)
Specification	human term placenta
Fixative:	Formalin
Antigen retrieval step:	Heat mediated - Buffer/Enzyme Used: 0.01M citrate buffer PH 6.0
Permeabilization:	No
Blocking step:	Serum as blocking agent for  2 hours  · Concentration: 1.5 % · Temperature: RT°C
Primary Antibody
Dilution	1/500
Incubation time:	18 hours  Temperature: 4°C Diluent: 1.5% normal goat serum in PBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: secondary antibody from Snta Cruz ABC Kit Conjugation: Biotin
Dilution:	1/200
Additional Data
Rating	Excellent
PubMed ID	PMID: 18986700: View
Additional notes	ab16030 staining SOCS3 in human term placenta showing strong staining within placental trophoblasts. Paraffin embedded human term placenta was incubated with ab16030 (1/500) at 4° overnight. Antigen retrieval was performed by heat induction in citrate buffer pH6.
Image	SOCS3 antibody for IHC-P (Human) Copyright image used under license

review by Antibody Solutions Ltd. submitted on 7 October 2008
Application:	Western blot
SOCS3 antibody (ab16030)
Sample	Mouse Cell lysate - whole cell (various: MEF, NIH-3T3, F9)
Loading amount:	30 µg
Specification	various: MEF, NIH-3T3, F9
Gel Running Conditions:	Reduced Denaturing (12.5% SDS PAGE)
Blocking step:	BSA as blocking agent for  2 hours  · Concentration: 3 % · Temperature: 21°C
Primary Antibody
Concentration	2 µg/ml
Incubation time:	12 hours  Temperature: 4°C Diluent: 3% BSA 0.1% Tween-20, PBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: Swine anti-Rabbit HRP Conjugation: HRP
Dilution:	1/5000
Detection
Detection method:	Supersignal West Dura
Exposure:	6 minutes and 0 seconds
Bands:	Specific: 28 kDa   Non-specific: 40, 50, 60 (faint) kDa
Additional Data
Rating	Good
Additional notes	Lane order (L-R): MEF, NIH-3T3, F9 This antibody detects a species of ~28kDa, which is consitent with the mobility of SOCS3 by SDS-PAGE. A few faint cross-reacting bands were observed at higher molecular weight, which could go away with diluting the antibody. I would recommend using BSA as the non-specific protein block with this antibody, as milk appeared to quench the specific signal (data not shown).
Image	SOCS3 antibody for WB (Mouse) Copyright image used under license

review by Carylyn Marek submitted on 8 April 2008
Application:	Flow Cytometry
SOCS3 antibody (ab16030)
Sample	Rat Cell (bone marrow-derived macrophages)
Specification	bone marrow-derived macrophages
Preparation:	Cell harvesting/tissue preparation method: Sample buffer:
Fixation:
Permeabilization:	Yes - BD fix and perm kit
Gating Strategy:
Primary Antibody
Dilution	1/300
Incubation time:	1 hour
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat anti-rabbit Conjugation: Alexa Fluor® 647
Dilution:	1/200
Additional Data
Rating	Excellent
Additional notes	Fixed and permeabilised using commercial fix and permeabilisation kit. Have tried several other SOCS3 antibodies from different companies and only this one works well for FACS. Picture: top is anti-rabbit conjugated alexa-fluor 647 secondary only. bottom is SOCS3 primary and anti-rabbit conjugated alexa-fluor 647 secondary together showing shift.
Image	SOCS3 antibody for Flow Cyt (Rat) Copyright image used under license