Anti-DCAMKL1 antibody ab31704 is a rabbit polyclonal antibody that is used in DCAMKL1 western blotting and immunofluorescence. Suitable for human, mouse and rat samples.
- Specificity confirmed with DCLK1 knockout cell line validation
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
ICC/IF | IP | WB | |
---|---|---|---|
Human | Predicted | Predicted | Tested |
Mouse | Predicted | Expected | Expected |
Rat | Expected | Predicted | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 0.5 µg/mL | Notes - |
Species Rat | Dilution info 0.5 µg/mL | Notes - |
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Probable kinase that may be involved in a calcium-signaling pathway controlling neuronal migration in the developing brain. May also participate in functions of the mature nervous system (By similarity).
Dcamkl1, Dclk, Dclk1, Dcamkl1, Dclk, Serine/threonine-protein kinase DCLK1, Doublecortin-like and CAM kinase-like 1, Doublecortin-like kinase 1
Anti-DCAMKL1 antibody ab31704 is a rabbit polyclonal antibody that is used in DCAMKL1 western blotting and immunofluorescence. Suitable for human, mouse and rat samples.
- Specificity confirmed with DCLK1 knockout cell line validation
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
Affinity purification Immunogen
This antibody recognizes 2 different human isoforms (AL: 82.2 KDa and BL: 47.6 KDa). From May 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help. You may also be interested in our alternative recombinant antibody, ab109029.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
ab31704 staining DCAMKL1 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab31704 at 5 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
DCAMKL1 was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to DCAMKL1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31704.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697).
Band: 82kDa: DCAMKL1; non specific - 52 and 27kDa: We are unsure as to the identity of this extra band.
All lanes: Immunoprecipitation - Anti-DCAMKL1 antibody (ab31704)
Predicted band size: 82 kDa
ICC/IF image of ab31704 stained PC12 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab31704, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
Lane 1: Wild type HAP1 whole cell lysate (20 μg)
Lane 2: DCAMKL1 knockout HAP1 whole cell lysate (20 μg)
Lane 3: NIH3T3 whole cell lysate (20 μg)
Lane 4: Human brain whole tissue lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab31704 observed at 90 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab31704 was shown to specifically react with DCAMKL1 in wild-type HAP1 cells along with additional cross reactive bands. No bands were observed when DCAMKL1 knockout cells were examined. Wild-type and DCAMKL1 knockout samples were subjected to SDS-PAGE. ab31704 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-DCAMKL1 antibody (ab31704)
Predicted band size: 82 kDa
The 82 kDa and 47 kDa bands correspond to the AL and BL isoforms respectively. This antibody should not detect the AS and BS isoforms of DCAMLK1.
All lanes: Western blot - Anti-DCAMKL1 antibody (ab31704) at 1 µg/mL
All lanes: Mouse Brain Whole Tissue Lysate at 20 µg
All lanes: IR Dye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
Performed under reducing conditions.
Predicted band size: 82 kDa
Observed band size: 47 kDa, 82 kDa
All lanes: Western blot - Anti-DCAMKL1 antibody (ab31704) at 1 µg/mL
All lanes: Brain (Rat) Tissue Lysate - normal tissue at 10 µg
All lanes: IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 82 kDa
Observed band size: 47 kDa, 82 kDa
The 82 kDa and 47 kDa bands correspond to the AL and BL isoforms respectively. DCAMLK1 has a number of potential phosphorylation sites which may explain the higher migrating bands at 52 and 54 kDa.
All lanes: Western blot - Anti-DCAMKL1 antibody (ab31704) at 1 µg/mL
All lanes: Human brain tissue lysate - total protein (ab29466) at 10 µg
All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 82 kDa
Observed band size: 30 kDa, 47 kDa, 52 kDa, 54 kDa, 82 kDa
Exposure time: 5min
All lanes: Western blot - Anti-DCAMKL1 antibody (ab31704) at 1 µg/mL
Lane 1: HCT116 WT at 30 µg
Lane 2: MiaPaCa2, at 30 µg
Lane 3: SW480 at 30 µg
Lane 4: SW620 at 30 µg
Lane 5: HT-29 at 30 µg
Lane 6: H4 at 30 µg
Lane 7: mouse brain lysate at 30 µg
All lanes: IRDye® 800CW Goat Anti-Rabbit, at 1/20000 dilution
Performed under reducing conditions.
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