ChromoTek Spot-tag® antibody (28A5)
Mouse monoclonal antibody to Spot-tag®
Host/IsoType
Mouse IgG1
Specificity
Spot-tag®
Applications
WB, ELISA, IF (ICC)
Conjugate
Unconjugated
CloneNo.
28a5
Cat no : 28a5
Synonyms
Validation Data Gallery
![Western blot analysis of N- and C-terminal Spot-tagged GFP added to HEK- 293T cell lysate. Detection with Spot-tag® antibody [28A5] (28a5, ChromoTek) 1:5,000 and Nano-Secondary® alpaca anti-mouse IgG1, recombinant VHH, Alexa Fluor® 488 [CTK0103, CTK0104] (sms1AF488-1, ChromoTek) 1:5,000. Western blot membrane was incubated simultaneously with the primary antibody and Nano-Secondary® (one-step staining).](https://www.ptglab.com/Products/Pictures/28A5-1.jpg "Western blot analysis of N- and C-terminal Spot-tagged GFP added to HEK- 293T cell lysate. Detection with Spot-tag® antibody [28A5] (28a5, ChromoTek) 1:5,000 and Nano-Secondary® alpaca anti-mouse IgG1, recombinant VHH, Alexa Fluor® 488 [CTK0103, CTK0104] (sms1AF488-1, ChromoTek) 1:5,000. Western blot membrane was incubated simultaneously with the primary antibody and Nano-Secondary® (one-step staining).")
![Western blot analysis of C-terminal Spot-tagged GFP added to HEK-293T cell lysate. Detection with Spot-tag® antibody [28A5] (28a5, ChromoTek) 1:5,000 and anti-mouse secondary antibody HRP 1:1,000.](https://www.ptglab.com/Products/Pictures/28A5-2.png "Western blot analysis of C-terminal Spot-tagged GFP added to HEK-293T cell lysate. Detection with Spot-tag® antibody [28A5] (28a5, ChromoTek) 1:5,000 and anti-mouse secondary antibody HRP 1:1,000.")
![Western blot analysis of different cell lysates +/- 50 ng C-terminal Spot-tagged GFP: human (HEK-293T), yeast (S.cerevisiae), insect (High Five™), bacteria (E.coli) and plant (A.thaliana). Detection with Spot-tag® antibody [28A5] (28a5, ChromoTek) 1:5,000 and Nano-Secondary® alpaca anti-mouse IgG1, recombinant VHH, Alexa Fluor® 488 [CTK0103, CTK0104] (sms1AF488-1, ChromoTek) 1:5,000. Western blot membrane was incubated simultaneously with the primary antibody and Nano-Secondary® (one-step staining).](https://www.ptglab.com/Products/Pictures/Result-28a5_20210803100622724.png "Western blot analysis of different cell lysates +/- 50 ng C-terminal Spot-tagged GFP: human (HEK-293T), yeast (S.cerevisiae), insect (High Five™), bacteria (E.coli) and plant (A.thaliana). Detection with Spot-tag® antibody [28A5] (28a5, ChromoTek) 1:5,000 and Nano-Secondary® alpaca anti-mouse IgG1, recombinant VHH, Alexa Fluor® 488 [CTK0103, CTK0104] (sms1AF488-1, ChromoTek) 1:5,000. Western blot membrane was incubated simultaneously with the primary antibody and Nano-Secondary® (one-step staining).")
![ELISA detection of N- and C-terminal Spot-tagged GFP with Spot-tag® antibody [28A5] (28a5, ChromoTek). N- and C-terminal Spot-tagged GFP was coated on a microtiter plate, blocked and detected with 1 nM Spot-tag® antibody [28A5] (28a5, ChromoTek) and anti-mouse IgG antibody AP.](https://www.ptglab.com/Products/Pictures/Result-28a5_20210803100844572.png "ELISA detection of N- and C-terminal Spot-tagged GFP with Spot-tag® antibody [28A5] (28a5, ChromoTek). N- and C-terminal Spot-tagged GFP was coated on a microtiter plate, blocked and detected with 1 nM Spot-tag® antibody [28A5] (28a5, ChromoTek) and anti-mouse IgG antibody AP.")
Product Information
Mouse monoclonal antibody to Spot-tag®
| Applications | WB, ELISA, IF (ICC) |
| Recommended Dilution | WB: 1:1,000 - 1:5,000 ELISA: 1:5,000 - 1:10,000 IF (ICC): 1:200 - 1:1,000 |
| Clonality | Monoclonal |
| Host/IsoType | Mouse IgG1 |
| Specificity/Target | Spot-tag® (PDRVRAVSHWSS) |
| RRID | AB_2892245 |
| Conjugate | Unconjugated |
| CloneNo. | 28a5 |
| Concentration | 1000 µg/mL |
| Storage Buffer | PBS, 50% glycerol, preservative: 0.09 % sodium azide |
| Storage Condition | Shipped on blue ice. Aliquot upon receipt and store at -20°C/-4°F. Avoid freeze-thaw cycles. |
Documentation
| SDS |
|---|
| 28a5 SDS Spot-tag® antibody [28A5] (EN) |
| Protocol |
|---|
| 28a5 Datasheet Spot-tag® antibody [28A5] |
Reviews
The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.
FH Tsimafei (Verified Customer) (06-01-2022) | mESCs expressed HA-SPOT-GFP protein (at a low level). IP was performed using in-house BC2 magnetic beads, followed by western blot. Membrane was probed with anti-SPOT antibodies at 4C, ON.
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FH Daniela (Verified Customer) (04-11-2022) | The 28A5 Spot-antibody was tested in Western blots of Expi-293F supernatants and purified proteins in a 1:5000 dilution using 5% non-fat milk (Biozym)/TBST (or 1x Rotiblock solution (Roth). Incubation was either 2 h at RT or ON at 4°C. Secondary HRP-antibody was incubated for 1 h at RT. The antibody is working fine, monomers, dimers, oligomers of the construct can be distinguished.
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