Immunofluorescence Customer Review for Anti-β-tubulin Antibody (STJ97037)
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Rating: Specificity: ANTIBODY CUSTOMER REVIEW Findings: “Some background staining seen on the negative control but very good staining otherwise.” – Marie Davies, The University of Sheffield Knowledge Dock Business Centre, University Way, London E16 2RD T: (+44) 0208 223 3081 F: (+44) 0207 681 2580 E: [email protected] w: www.stjohnslabs.com Protocol Tissue Processing When cells were cultured to 70%-80% plating density on coverslips, cells were fixed in 4% PFA for 20 mins at room temperature before application of antibodies and plating coverslips on slides with DAPI Fluoromount. Primary Antibody Probing Primary antibody was added to the blocking solution at 1:100 for 1hr room temperature. Fixing 4% PFA in PBS 20 mins room temperature Washing 1X PBS wash for 3 times. Permeabilization 1X PBS 0.4% Triton-X-100 15 mins room temperature Secondary Antibody Probing Secondary antibody was diluted 1:500 and DAPI was diluted 1:1000 in 1% bovine serum albumin in PBS and incubated for 1 hr at room temperature. Washing 1X PBS wash for 3 times. Washing 1X PBS wash for 3 times Blocking 1X PBS 0.01% Triton-X-100 0.2% fish skin gelatin 1hr room temperature Mounting Mounted coverslips on slides with DAPI fluoromount. Observed on Confocal Microscopy Model Number STJ97037 Antibody Name Anti-β-tubulin antibody Application Immunofluorescence Species Human Tissue NSC-34 Cells Dilution 1:100 Secondary Antibody Anti-mouse IgG Alexa Fluor 488 Secondary Dilution 1:2000 Fig. In NSC-34 cell line, there was detection of B-tubulin by STJ97037 Actin-B-tubulin Antibody (Figure: Left : No Primary Antibody Staining, Right: STJ97037 Actin-B-tubulin Antibody 1:100)
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Transcript of Immunofluorescence Customer Review for Anti-β-tubulin Antibody (STJ97037)
Rating:
Specificity:
ANTIBODY CUSTOMER REVIEWFindings: “Some background staining seen on the negative control but very good staining otherwise.” – Marie Davies, The University of Sheffield
Knowledge Dock Business Centre, University Way, London E16 2RD T: (+44) 0208 223 3081 F: (+44) 0207 681 2580 E: [email protected] w: www.stjohnslabs.com
Protocol
Tissue Processing When cells were cultured to 70%-80% plating density on coverslips, cells were fixed in 4% PFA for 20 mins at room temperature before application of antibodies and plating coverslips on slides with DAPI Fluoromount.
Primary Antibody
Probing
Primary antibody was added to the blocking solution at 1:100 for 1hr room temperature.
Fixing 4% PFA in PBS 20 mins room temperature
Washing 1X PBS wash for 3 times.
Permeabilization 1X PBS 0.4% Triton-X-100 15 mins room temperature
Secondary Antibody
Probing
Secondary antibody was diluted 1:500 and DAPI was diluted 1:1000 in 1% bovine serum albumin in PBS and incubated for 1 hr at room temperature.
Washing 1X PBS wash for 3 times. Washing 1X PBS wash for 3 times
Blocking 1X PBS 0.01% Triton-X-100 0.2% fish skin gelatin 1hr room temperature
Mounting Mounted coverslips on slides with DAPI fluoromount. Observed on Confocal Microscopy
Model Number STJ97037
Antibody Name Anti-β-tubulin antibody
Application Immunofluorescence
Species Human
Tissue NSC-34 Cells
Dilution 1:100
Secondary Antibody Anti-mouse IgG Alexa Fluor 488
Secondary Dilution 1:2000
Fig. In NSC-34 cell line, there was detection of B-tubulin by STJ97037 Actin-B-tubulin Antibody (Figure: Left : No Primary Antibody Staining, Right: STJ97037 Actin-B-tubulin Antibody 1:100)
