WDR34 Antibody - BSA Free

Western Blot: WDR34 Antibody [NBP1-88805] - Sucrose density gradient centrifugation of cell lysates from RPE1 cells. RPE1 cells were lysed and loaded onto a 5-40% sucrose density gradient. After centrifugation, 1-ml fractions were removed from the gradient and probed by immunoblotting to determine the distribution across the gradient of DIC74 (DYNC1I2), WDR34 or GAPDH. Image collected and cropped by CiteAb from the following publication (//jcs.biologists.org/cgi/doi/10.1242/jcs.159038), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: WDR34 Antibody [NBP1-88805] - Localization of mGFP-WDR34. Cells expressing mGFP-WDR34 were fixed with paraformaldehyde and labeled to detect acetylated tubulin and WDR34, as indicated. Two examples are shown. Cells expressing GFP were processed as in E to demonstrate an absence of GFP from primary cilia. Scale bars: 5 um. Image collected and cropped by CiteAb from the following publication (//jcs.biologists.org/cgi/doi/10.1242/jcs.159038), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: WDR34 Antibody [NBP1-88805] - Staining of human stomach shows strong cytoplasmic positivity in glandular cells.

Western Blot: WDR34 Antibody [NBP1-88805] - The stabilities of WDR34 and WDR60 are interdependent. Immunoblotting confirmed the efficacy of WDR34 and WDR60 siRNAs. Immunoblotting for WDR34 following suppression of WDR60 and vice versa was also used to test the interdependency of these subunits. Lamin A/C is included as a negative control, GAPDH as a loading control. Molecular-mass markers are indicated (kDa). Image collected and cropped by CiteAb from the following publication (//jcs.biologists.org/cgi/doi/10.1242/jcs.159038), licensed under a CC-BY license.

Knockout Validated: WDR34 Antibody [NBP1-88805] - HA-WDR34 (NBP1-88805) and HA-WDR60 (NBP1-90437) expression in WT and KO cells. (A) Pull down of HA-WDR60 in WT and WDR34 KO cells. HA-WDR60 pulls down WDR34 in WT but not in KO cells (B) Pull down of HA-WDR34 in WT and WDR60 KO cells. HA-WDR34 pulls down WDR60 in WT but not in KO cells. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547/) licensed under a CC-BY license.

Western Blot: WDR34 Antibody [NBP1-88805] - Immunoprecipitation from serum-starved RPE1 cells stably expressing either mGFP or mGFP–WDR34. GFP-traps of cells expressing mGFP alone or mGFP–WDR34 were separated by SDS-PAGE & immunoblotted to detect (A) WDR34, (B) WDR60, (C) IC74 (DYNC1I2), (D) GAPDH, (E) p150Glued, (F) p50dynamitin, (G) LIS1, (H) Tctex-1, (I) NudCD3 or (J) TCTEX1D2, as indicated. Molecular-mass markers are shown (kDa). In each case, the lanes show the input (‘I’), the unbound fraction (‘U’) & the bound fraction (‘B’), as indicated. The asterisk in J indicates non-specific bands. Image collected & cropped by CiteAb from the following publication (//journals.biologists.com/jcs/article/doi/10.1242/jcs.159038/259056/Subunit-composition-of-the-human-cytoplasmic), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] - (A) RNAi of WDR34 & WDR60 validated by immunoblotting with GAPODH as a loading control.(B) Immunofluorescence of TMEM67 & RPGRIP1L in WDR34 & WDR60 depleted cells. TMEM67 is seen at the base & within the cilium proximal to this in both control & depleted cells, RPGRIP1L is more tightly restricted to the base of the cilium in all cases. These are representative images, we have analysed multiple fields of view in each case using z-stacks & cannot identify any difference. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] - CRISPR control cells lines show no defect in ciliogenesis.(A & B) Immunoblotting for WDR34 & WDR60 in WT & KO cells. (A) A 52 kDa band, corresponding to WDR34 is lost in WDR34 KO cells lysate. (B) A WDR60 band of 123 kDa is lost in the WDR60 KO cells lysate. (C) WDR34 KO CTRL & WDR60 KO CTRL cells stained with Arl13b (green) & AcTub (red). (D) WDR60 KO CTRL cells stained with IFT88 (green). (Di) IFT88 intensity quantification in WT & WDR60 KO CTRL cells (n = 3, 98 WT, 102 WDR60 KO CTRL, & 122 WDR60 KO cells quantified). Mann-Whitney test was used, p-value: *=<0.05 as above. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] - HA-WDR34 & HA-WDR60 expression in WT & KO cells.(A) Pull down of HA-WDR60 in WT & WDR34 KO cells. HA-WDR60 pulls down WDR34 in WT but not in KO cells (B) Pull down of HA-WDR34 in WT & WDR60 KO cells. HA-WDR34 pulls down WDR60 in WT but not in KO cells. (C) Immunoprecipitation of HA-tagged GFP, WDR60, & WDR34 followed by immunoblot for LIC3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] - HA-WDR34 & HA-WDR60 expression in WT & KO cells.(A) Pull down of HA-WDR60 in WT & WDR34 KO cells. HA-WDR60 pulls down WDR34 in WT but not in KO cells (B) Pull down of HA-WDR34 in WT & WDR60 KO cells. HA-WDR34 pulls down WDR60 in WT but not in KO cells. (C) Immunoprecipitation of HA-tagged GFP, WDR60, & WDR34 followed by immunoblot for LIC3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] - Dynein-2 assembly in primary cilium.(A) Immunoblotting for WDR60 & WDR34 in WT, WDR34 KO#1 & WDR60 KO cells. Arrows indicate WDR34 & WDR60 proteins. (B) LIC3/DYNC2LI1 localization in the cilia of WT, WDR34 KO#1 & WDR60 KO cells. (C) DHC2/DYNC2H1 localization at the ciliary base in WT & KO cells. (Ci) Intensity quantification shows a reduction of DHC2/DYNC2H1 at the ciliary base in WDR34 KO#1 cells (n = 3, 120 WT, 106 WDR60 KO, & 71 WDR34 KO #1 cells quantified). (D) TCTEX1/DYNLT1 localizes at the ciliary base in WT & KO cells. (Di) Intensity quantification of TCTEX1/DYNLT1 at the ciliary base (n = 3 115 WT, 85 WDR60 KO, & 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5 μm. Arrows point to the ciliary base.Overexpression of WDR34 cannot rescue WDR60 KO phenotype & vice versa.(A) HA-WDR60 localization in WT & WDR34 KO cells. (Ai) Intensity quantification of HA-WDR60 in primary cilia (n = 3, 50 WT & 50 WDR34 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (B) HA-WDR34 localization in WT & WDR60 cells. For HA immunolabeling in Fig. A & B cells were treated with cytoskeletal buffer as described in methods. (C) Arl13b staining of WDR34 KO cells expressing HA-WDR60. HA labeling in green shows stable expression of HA-WDR60. (D) IFT88 localizes predominantly at the ciliary base in WT cells whereas WDR60 KO cells stably expressing HA-WDR34 accumulate IFT88 at the ciliary tip, similar to untransfected WDR60 KO cells. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] - Dynein-2 assembly in primary cilium.(A) Immunoblotting for WDR60 & WDR34 in WT, WDR34 KO#1 & WDR60 KO cells. Arrows indicate WDR34 & WDR60 proteins. (B) LIC3/DYNC2LI1 localization in the cilia of WT, WDR34 KO#1 & WDR60 KO cells. (C) DHC2/DYNC2H1 localization at the ciliary base in WT & KO cells. (Ci) Intensity quantification shows a reduction of DHC2/DYNC2H1 at the ciliary base in WDR34 KO#1 cells (n = 3, 120 WT, 106 WDR60 KO, & 71 WDR34 KO #1 cells quantified). (D) TCTEX1/DYNLT1 localizes at the ciliary base in WT & KO cells. (Di) Intensity quantification of TCTEX1/DYNLT1 at the ciliary base (n = 3 115 WT, 85 WDR60 KO, & 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5 μm. Arrows point to the ciliary base.Overexpression of WDR34 cannot rescue WDR60 KO phenotype & vice versa.(A) HA-WDR60 localization in WT & WDR34 KO cells. (Ai) Intensity quantification of HA-WDR60 in primary cilia (n = 3, 50 WT & 50 WDR34 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (B) HA-WDR34 localization in WT & WDR60 cells. For HA immunolabeling in Fig. A & B cells were treated with cytoskeletal buffer as described in methods. (C) Arl13b staining of WDR34 KO cells expressing HA-WDR60. HA labeling in green shows stable expression of HA-WDR60. (D) IFT88 localizes predominantly at the ciliary base in WT cells whereas WDR60 KO cells stably expressing HA-WDR34 accumulate IFT88 at the ciliary tip, similar to untransfected WDR60 KO cells. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] - Dynein-2 assembly in primary cilium.(A) Immunoblotting for WDR60 & WDR34 in WT, WDR34 KO#1 & WDR60 KO cells. Arrows indicate WDR34 & WDR60 proteins. (B) LIC3/DYNC2LI1 localization in the cilia of WT, WDR34 KO#1 & WDR60 KO cells. (C) DHC2/DYNC2H1 localization at the ciliary base in WT & KO cells. (Ci) Intensity quantification shows a reduction of DHC2/DYNC2H1 at the ciliary base in WDR34 KO#1 cells (n = 3, 120 WT, 106 WDR60 KO, & 71 WDR34 KO #1 cells quantified). (D) TCTEX1/DYNLT1 localizes at the ciliary base in WT & KO cells. (Di) Intensity quantification of TCTEX1/DYNLT1 at the ciliary base (n = 3 115 WT, 85 WDR60 KO, & 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5 μm. Arrows point to the ciliary base.Overexpression of WDR34 cannot rescue WDR60 KO phenotype & vice versa.(A) HA-WDR60 localization in WT & WDR34 KO cells. (Ai) Intensity quantification of HA-WDR60 in primary cilia (n = 3, 50 WT & 50 WDR34 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (B) HA-WDR34 localization in WT & WDR60 cells. For HA immunolabeling in Fig. A & B cells were treated with cytoskeletal buffer as described in methods. (C) Arl13b staining of WDR34 KO cells expressing HA-WDR60. HA labeling in green shows stable expression of HA-WDR60. (D) IFT88 localizes predominantly at the ciliary base in WT cells whereas WDR60 KO cells stably expressing HA-WDR34 accumulate IFT88 at the ciliary tip, similar to untransfected WDR60 KO cells. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] - Dynein-2 assembly in primary cilium.(A) Immunoblotting for WDR60 & WDR34 in WT, WDR34 KO#1 & WDR60 KO cells. Arrows indicate WDR34 & WDR60 proteins. (B) LIC3/DYNC2LI1 localization in the cilia of WT, WDR34 KO#1 & WDR60 KO cells. (C) DHC2/DYNC2H1 localization at the ciliary base in WT & KO cells. (Ci) Intensity quantification shows a reduction of DHC2/DYNC2H1 at the ciliary base in WDR34 KO#1 cells (n = 3, 120 WT, 106 WDR60 KO, & 71 WDR34 KO #1 cells quantified). (D) TCTEX1/DYNLT1 localizes at the ciliary base in WT & KO cells. (Di) Intensity quantification of TCTEX1/DYNLT1 at the ciliary base (n = 3 115 WT, 85 WDR60 KO, & 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5 μm. Arrows point to the ciliary base.Overexpression of WDR34 cannot rescue WDR60 KO phenotype & vice versa.(A) HA-WDR60 localization in WT & WDR34 KO cells. (Ai) Intensity quantification of HA-WDR60 in primary cilia (n = 3, 50 WT & 50 WDR34 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (B) HA-WDR34 localization in WT & WDR60 cells. For HA immunolabeling in Fig. A & B cells were treated with cytoskeletal buffer as described in methods. (C) Arl13b staining of WDR34 KO cells expressing HA-WDR60. HA labeling in green shows stable expression of HA-WDR60. (D) IFT88 localizes predominantly at the ciliary base in WT cells whereas WDR60 KO cells stably expressing HA-WDR34 accumulate IFT88 at the ciliary tip, similar to untransfected WDR60 KO cells. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF, IHC, IP, KO, Mycoplasma
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

WDR34 Antibody - BSA Free Summary

• Description: Novus Biologicals Knockout (KO) Validated Rabbit WDR34 Antibody - BSA Free (NBP1-88805) is a polyclonal antibody validated for use in IHC, WB, ICC/IF and IP. Anti-WDR34 Antibody: Cited in 5 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: This antibody was developed against Recombinant Protein corresponding to amino acids: APPLTSLQLSLKYLFAVRWSPVRPLVFAAASGKGDVQLFDLQKSSQKPTVLIKQTQDESPVYCLEFNSQQTQLLAAGDAQGTVKV
• Predicted Species: Mouse (91%), Rat (91%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: WDR34
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence Reported in the scientific literature (PMID: 25205765)
  • Immunohistochemistry 1:50 - 1:200
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Immunoprecipitation Reported in scientific literature (PMID:30320547)
  • Knockdown Validated
  • Knockout Validated
  • Western Blot Reported in the scientific literature (PMID: 25205765)
• Application Notes: For IHC-Paraffin, HIER pH 6 retrieval is recommended.
• Control Peptide:

  WDR34 Protein (NBP1-88805PEP)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH 7.2) and 40% Glycerol
• Preservative: 0.02% Sodium Azide
• Purity: Immunogen affinity purified

Alternate Names for WDR34 Antibody - BSA Free

• bA216B9.3
• MGC20486
• WD repeat domain 34
• WD repeat-containing protein 34

Background

WDR34 encodes a member of the WD repeat protein family. WD repeats are minimally conserved regions of approximately 40 amino acids typically bracketed by gly-his and trp-asp (GH-WD), which may facilitate formation of heterotrimeric or multiprotein complexes. Members of this family are involved in a variety of cellular processes, including cell cycle progression, signal transduction, apoptosis, and gene regulation. [provided by RefSeq]

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for WDR34 Antibody (NBP1-88805)(6)

Publications using NBP1-88805

• Weijman J, Vuolo L, Shak C et al. Roles for CEP170 in cilia function and dynein-2 assembly bioRxiv 2023-11-21 (WB, Human)
• Weijman JF, Vuolo L, Shak C et Al. Roles for CEP170 in cilia function and dynein-2 assembly J Cell Sci 2024-05-01 [PMID: 38533689]
• Vuolo Laura, Stevenson Nicola L, Heesom Kate J, Stephens David J Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function. International Immunology 2018-10-16 [PMID: 30320547] (KD, WB, IP, Human)
• Yamamoto JI, Kasamatsu A, Okubo Y et al. Evaluation of tryptophan-aspartic acid repeat-containing protein 34 as a novel tumor-suppressor molecule in human oral cancer. Biochem. Biophys. Res. Commun. 2017-12-23 [PMID: 29278705] (Human)
• Asante D, Stevenson NL, Stephens DJ. Subunit composition of the human cytoplasmic dynein-2 complex. J Cell Sci 2014-11-01 [PMID: 25205765] (ICC/IF, WB, Human)
• Asante D, MacCarthy-Morrogh L, Townley AK et al. A role for the Golgi matrix protein giantin in ciliogenesis through control of the localization of dynein-2. J Cell Sci 2013-11-15 [PMID: 24046448] (WB, Human)

Bioinformatics

• Gene Symbol: WDR34