Punches were blocked in IF buffer (1% BSA and 0.05% Tween‐20 in PBS) for 1–2 h at RT, followed by the incubation with primary antibody diluted in IF buffer, for 24 h at 4°C. Punches were washed in 1× PBS for 1–2 h and incubated for 24 h at 4°C with secondary antibody and DAPI (ThermoFisher; D1306) diluted 1:10 000 in IF buffer. The following primary antibodies were used for immunostaining of expanded samples: mouse anti‐acetylated tubulin (Sigma, T7451) at 1:4000, mouse anti‐alpha tubulin (Sigma; T6074) at 1:500, rabbit antibeta tubulin (Abcam; ab15568) at 1:250, rabbit anti‐ARL13B (Abcam; ab83879) at 1:150, rabbit anti‐Cep164 (Proteintech; 22227‐1‐AP) at 1:500, rabbit anti‐ANKRD26 (GeneTex; GTX128255) at 1:200, rabbit anti‐FBF1 (sigma; HPA023677) at 1:100, rabbit anti‐RPGRIP1L (Proteintech; 55160‐1‐AP) at 1:150, rabbit anti‐IFT88 (Proteintech; 13967‐1‐AP) at 1:150, mouse anti‐Rootletin (Santa Cruz; sc‐374056) at 1:50, rabbit antipericentrin (Abcam; ab4448) at 1:400, rabbit anti‐Cep290 (Abcam; ab84870) at 1:150 and rabbit antipolyglutamylated tubulin (AdipoGen; AG‐25B‐0030) at 1:800. Secondary antibodies antimouse Alexa Fluor 488 (Invitrogen; A11029), antirabbit Alexa Fluor 488 (Invitrogen; A11034), antimouse Alexa Fluor 555 (Invitrogen; A28180), antirabbit Alexa Fluor 555 (Invitrogen; A21429) were used at a 1:800 dilution to label primary antibodies.
Ab84870
Manufactured by Abcam
Ab84870 is a laboratory antibody product from Abcam. It is a recombinant polyclonal antibody.
Automatically generated - may contain errors
Lab products found in correlation
T7451, by Merck Group (2 mentions)
D1306, by Thermo Fisher Scientific (1 mentions)
A11029, by Thermo Fisher Scientific (1 mentions)
A11034, by Thermo Fisher Scientific (1 mentions)
A21429, by Thermo Fisher Scientific (1 mentions)
T6074, by Merck Group (1 mentions)
Ag 25b 0030, by Adipogen (1 mentions)
A28180, by Thermo Fisher Scientific (1 mentions)
Ab15568, by Abcam (1 mentions)
Ab4448, by Abcam (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa buffer, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Cep290, by Abcam (1 mentions)
Sab3500022, by Merck Group (1 mentions)
β actin sc 47778, by Santa Cruz Biotechnology (1 mentions)
Alx 804 885 c100, by Enzo Life Sciences (1 mentions)
Talpid3, by Proteintech (1 mentions)
Ift140, by Proteintech (1 mentions)
3 protocols using ab84870
1
Immunostaining of Expanded Samples
2
Ciliary Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293 cells and 2 dpf zebrafish embryos were homogenized in RIPA buffer (Cell Signaling Technology, 9806) containing Protease Inhibitor Cocktail (Roche, 04693159001) followed by short sonication. Sample supernatants were collected after centrifugation at 18,400 g for 20 min. Protein (30 μg) was separated on a NuPAGE™ Mini Protein Gel (Invitrogen, NP0322BOX) before being transferred to PVDF membrane (Millipore, IPVH00010). Membranes were blocked in TBS containing 5% non-fat milk powder (Lab Scientific) for 1 h. Immunoblotting was performed at 4°C overnight using antibodies against TMEM67 (1:1000; 13975-1-AP), TCTN1 (1:1000; 15004-1-AP), B9D2 (1:1000; 22508-1-AP) and MKS5/RPGRIP1L (1:1000; 29778-1-AP) from Proteintech; NPHP1 (1:1000; GTX65891) from GeneTex; IFT172 (1:1000; bs-15560R) from Thermo Fisher Scientific; NPHP4 (1:1000; H00261734-M01A) from Abnova; CEP290 (1:1000; ab84870) from Abcam; and GAPDH (1:2000; 2118) from Cell Signaling Technology. Antibody against Ift88 (1:3000) was generously gifted by Dr Brian Perkins (Cole Eye Institute, Cleveland Clinic, Cleveland, OH, USA). The membranes were incubated with a secondary antibody at room temperature for 1 h after washing with TBS containing 0.5% Tween 20. Protein bands were visualized using Pierce Clarity Western ECL substrate (Bio-Rad, 170-5061).
3
Antibodies for Ciliogenesis and Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following primary antibodies were used: acetylated α-tubulin (T7451, diluted 1:2000 for immunofluorescence, Sigma), γ-tubulin (T6557, Sigma), FLAG (F1804, Sigma), HA (H3663, Sigma), CEP83 (HPA038161, Sigma), CEP164 (SAB3500022, Sigma), CEP290 (ab84870, Abcam), TALPID3 (24421-1-AP, Proteintech), ARL13b (17711-1-AP, Proteintech), FBF1 (11531-1-AP, Proteintech), TCTN1 (15004-1-AP, Proteintech), SCLT1 (14875-1-AP, Proteintech), IFT140 (17460-1-AP, Proteintech), β-actin (sc-47778, diluted 1:5000 for western blotting, Santa Cruz), SMO (sc-166685, Santa Cruz), CEP89 (ab204410, Abcam), ANKRD26 (GTX128255, GeneTex), polyglutamylated tubulin (ALX-804-885-C100, diluted 1:2000 for immunofluorescence, Enzo Life Sciences), GLI-3 (AF3690, R&D Systems), ODF2 (H00004957-M01, Abnova), and Polycystin-2 (Baltimore Polycystic Kidney Disease (PKD) Research and Clinical Core Center). The secondary antibodies were goat anti-mouse Alexa Fluor 488/594 or goat anti-rabbit Alexa Fluor 488/594 (1:1000 dilution). Primary antibodies were diluted 1:500 for immunofluorescence and 1:2000 for western blotting experiments unless otherwise specified. Uncropped versions of blots can be found in Supplementary Fig. 9 .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!